Thermo Scientific Pierce_dialysishandbook [m34mr7x7wzl6]

Thermo Scientific Pierce_dialysishandbook [m34mr7x7wzl6]
June 14, 2021 0 Comments

Thermo Scientific Pierce® Excessive-Efficiency Dialysis and Desalting Technical Handbook That includes Thermo Scientific Slide-A-Lyzer® Dialysis Cassettes

Excessive-Efficiency Dialysis Product Choice Information

10-100 µl

0.1-30 ml

15-100 ml

Thermo Scientific Slide-A-Lyzer® MINI Dialysis Unit

Thermo Scientific Slide-A-Lyzer Dialysis Cassette

Thermo Scientific SnakeSkin® Dialysis Tubing

MWCO Membrane

Web page 4

Web page 5

Web page 6

2K

N/A

X

N/A

3.5K

X

X

X

7K

X

X

X

10K

X

X

X

20K

N/A

X

N/A

Desk of Contents Excessive-Efficiency Dialysis

1

New! RED Speedy Equilibrium Dialysis Gadget

Dialysis: An Overview

2

Slide-A-Lyzer Concentrating Answer

14

Continuously Requested Questions

3

14

Slide-A-Lyzer MINI Dialysis Items

4

Concentrating with Slide-A-Lyzer MINI Dialysis Items

Slide-A-Lyzer Dialysis Cassettes

5

Microdialyzer System

15

SnakeSkin Dialysis Tubing

6

Pressured Dialysis for Pattern Focus

16

2K MWCO Membrane Merchandise

7

Evaporation for Pattern Focus

16

3.5K MWCO Membrane Merchandise

8

7K MWCO Membrane Merchandise

9

10K MWCO Membrane Merchandise

10

New! 20K MWCO Membrane Merchandise

11

12-13

Desalting Columns and Plates Zeba™ Desalt Spin Columns New! Zeba 96-Effectively Desalt Spin Plates D-Salt Columns

17-19 17 18 19

Cell Lysis Options

20-21

Excessive-Efficiency Dialysis Dialysis is a separation method that gained recognition in life science laboratories throughout the Fifties. Analysis papers of that period described dialysis as a brand new, cutting-edge software that scientists might use to unravel advanced mixtures of biomacromolecules. Lots of the dialysis theories established at the moment are the cornerstones for up to date merchandise featured on this brochure. There are, nonetheless, two main variations between the dialysis instruments of yesterday and as we speak – preparation time and the quantity of pattern loss attributable to leaks. Early laboratory dialysis strategies concerned dedicating a significant period of time to membrane

Previous dialysate discarded and changed with 1,000 ml of 100 mM PBS, pH 7.6

2 hours

3 ml of 1 mg/ml IgG in 0.1 M Tris buffer, pH 7.4 inside a Slide-A-Lyzer Dialysis Cassette (10K MWCO) positioned in 1,000 ml of 100 mM PBS, pH 7.6.

= Tris

New developments in dialysis strategies had been stagnant throughout the previous few many years, whereas ultrafiltration methods flourished fueled by advances in non-cellulose membranes and accessibility of bench-top centrifuges. Ultrafiltration through centrifugation was the established conference till we launched the Slide-A-Lyzer Dialysis Cassette in 1994. See a product demonstration at www.thermo.com/dialysis

Enlarged view of Slide-A-Lyzer Cassette inside

Enlarged view of Slide-A-Lyzer Cassette inside

= PBS

preparation; Thermo Scientific Pierce Dialysis Merchandise are primarily prepared to make use of and resist pattern leakage.

= IgG

Enlarged view of Slide-A-Lyzer Cassette inside

Previous dialysate discarded and changed with 1,000 ml of 100 mM PBS, pH 7.6

2 hours

IgG would diffuse out if it might, however it’s too massive to enter the pores within the membrane; due to this fact, the quantity of IgG contained in the Cassette stays fixed. The Tris buffer focus drops under 0.01 M contained in the Cassette because the Tris buffer diffuses out and the PBS buffer diffuses in.

IgG contained in the Cassette stays fixed. The Tris buffer contained in the Cassette drops to close undetectable ranges. The buffer contained in the Cassette is 100 mM PBS, pH 7.6.

To order, name 800-874-3723 or 815-968-0747. Exterior america, contact your native department workplace or distributor.

1

Dialysis: An Overview Dialysis is the separation of small and enormous molecules in an answer by selective diffusion by a semipermeable membrane. Sometimes a pattern containing a protein or nucleic acid will comprise undesirable small molecular weight (MW) compounds reminiscent of a buffer salt (Tris, PBS, and so on.), a lowering agent [dithiothreitol (DTT), β-mercaptoethanol (BME), etc.] or a preservative (sodium azide, thimerosol, and so on.). The pattern is contained contained in the dialysis membrane. A dialysate 200 to 300 instances the quantity of the pattern is outdoors the dialysis 0.5-3 ml membrane, which creates and maintains a focus differential throughout the membrane. As soon as the liquid-to-liquid interface (pattern on one facet of the membrane and dialysate on the opposite) is initiated, all molecules will attempt to diffuse in both course throughout the membrane to succeed in equilibrium. Dialysis (diffusion) will cease when equilibrium is achieved. Usually the speed of dialysis slows as equilibrium approaches, requiring the dialysate be modified after a number of hours to re-create Fluid Chamber the focus differential that drives the • dialysis course of.

“O” Rings

The membrane is the important thing to dialysis. The • • semipermeable membrane accommodates pores of a recognized dimension vary that Filling Ports are massive sufficient to let • small MW compounds cross by, however prohibit massive MW compounds (e.g., proteins and nucleic To Rotating Motor acids). The best membrane • is skinny, has quite a few pores of uniform diameter, and doesn’t bind proteins and nucleic acids. What scientists have been utilizing for many years is an extruded regenerated cellulose membrane that’s near a great membrane. Take a look at Membrane

12-30 ml

0.1-0.5 ml

3-12 ml

10-100 µl Thermo Scientific Slide-A-Lyzer MINI Dialysis Unit

Different essential variables are pattern and dialysate quantity. The best state of affairs is to have a small pattern quantity and a big dialysate quantity to maximise the focus differential. The pattern quantity is essential as a result of subsequent functions have sure minimal quantity necessities. Nevertheless, after the minimal quantity necessities are met, it isn’t advantageous to dialyze extra pattern than is required. Relying on the floor space of a given pattern, a small quantity pattern will dialyze a lot quicker than a big quantity pattern. Not solely is expending extra time wasteful, it may end up in pattern loss as a result of the longer a pattern is in touch with solid-phase surfaces, the extra seemingly proteins or nucleic acids will nonspecifically bind or denature. Reference 1. Klein, E., et al. DHEW Publication No. 77-1294, p. 17.

IgG (160 Å)

Pore (24 Å)

Nevertheless, most scientists usually assume an excessive amount of chromatographic decision related to the membrane’s molecular weight cutoff (MWCO). We decide the MWCO of our dialysis membrane through the use of the rotating batch dialysis cell (see diagram1 above). Within the rotating cell, the membrane to be examined is held in place between two round cavities of equal dimension. One facet of the cell is partially full of an answer containing a molecule of recognized MW. The opposite facet is full of an equal quantity of buffer or saline. The options are combined and saved in touch with the membrane by rotating the cell at a relentless pace. The MW customary focus in every half of the cell is measured after a set time frame and the p.c retention is calculated. This kind of system gives a extra correct MWCO willpower than utilizing ultrafiltration strategies that measure hydraulic permeability or volumetric flux vs. stress utilizing saline or buffer alone.

2

15-100 ml Thermo Scientific SnakeSkin Dialysis Tubing

10K Membrane

2-ME (5 Å)

Determine 1. Diffusion of particles by a semi-permeable membrane.

For extra data, or to obtain a product instruction booklet, go to www.thermo.com/pierce

Continuously Requested Questions About Dialysis 1) How exact is the MWCO? The MWCO is reproducible, however not very exact. When selecting which MWCO membrane to make use of, it’s advisable to have each the excessive MW compounds that you just need to retain, and the low MW compounds that you just need to diffuse out as far faraway from the membrane’s MWCO as potential. Our dialysis merchandise can be found with 2K, 3.5K, 7K, 10K and 20K MWCO membranes. The retention profile exhibited is clearly distinct and reproducible for every MWCO membrane when testing compounds of recognized MW. We don’t promote merchandise with regenerated cellulose membranes with MWCOs under 2K and above 20K as a result of they can’t be manufactured to our high-quality requirements at the moment.

2) Is stirring mandatory? Stirring considerably decreases the dialysis time. All membranes possess an inside pores and skin, which specialists have described as “seaweed-like,” and an outer pores and skin. There aren’t any channels of a set diameter extending from the pattern facet by to the dialysate facet. As an alternative, low MW compounds from the pattern diffuse into the inside pores and skin pores then by the membrane inside. These low MW compounds exit by a pore within the outer pores and skin of the membrane, to a micro-environment known as the Nernst layer. On this layer, which is roughly 200-300 molecules thick, low MW compounds are at a better focus in relation to the remainder of the dialysate. Stirring, which effectively breaks up the macro-environment outdoors the Nernst layer, shortly restores the focus differential wanted to drive the diffusion course of.

3) Is temperature essential? Temperature is considerably essential as a result of molecules transfer and diffuse quicker at increased temperatures; nonetheless, sustaining the viability of your pattern is the precedence. So the standard vary for dialysis is from ambient to cold-room temperatures.

4) When is my dialysis finished? There isn’t a simply measured dialysis endpoint. The purpose is to cut back the focus of low MW compounds to a stage that won’t intervene with subsequent steps in your experiment. Customary follow has been as follows: 1) Dialyze for two hours at room temperature (RT), 2) Change the dialysate earlier than dialyzing for one more 2 hours at RT, and three) Change the dialysate once more and dialyze for 1 hour to in a single day within the chilly room. Thermo Scientific Pierce Excessive-Efficiency Dialysis Merchandise make the dialysis course of quicker than ever. The essential precept of the Slide-A-Lyzer MINI Dialysis Unit is to deposit a ten µl pattern (primarily a monolayer) on a dialysis membrane in touch with a dialysate that’s 100,000 instances bigger than the pattern quantity. Small MW compounds have an especially brief (

5) Is membrane pretreatment mandatory? A brief hydration is critical for some MWCO membranes within the Slide-A-Lyzer Dialysis Cassette product line. In any other case, the regenerated cellulose membranes are clear and require no pretreatment. A really small quantity of both glycerine or sulfur could also be current. These low MW compounds will diffuse out of the membrane and into the dialysate throughout the regular dialysis course of. If mandatory, these compounds could also be dialyzed forward of time however that is often pointless.

6) When pattern is injected into the Slide-A-Lyzer Dialysis Cassette, the membrane generally folds. What causes this? As a result of the dialysis membrane is manufactured as a tube, the regenerated cellulose polymer has “reminiscence” and needs to return to that form regardless that the tube was minimize right into a flat membrane. Due to this fact, when a membrane is hydrated and the Cassette is stuffed, the membrane will stretch or pull otherwise with respect to the X-axis or Y-axis. Though this does have minor implications relative to floor space, these Slide-A-Lyzer Dialysis Cassettes will operate simply superb.

To order, name 800-874-3723 or 815-968-0747. Exterior america, contact your native department workplace or distributor.

3

Thermo Scientific Slide-A-Lyzer MINI Dialysis Items For pattern volumes as small as 10 μl

The Slide-A-Lyzer MINI Dialysis Unit is a small disposable cup manufactured from polypropylene and regenerated cellulose. Pattern is added and eliminated simply utilizing an ordinary laboratory pipette. A float (offered individually) holds the Slide-A-Lyzer MINI Dialysis Unit upright, floating on the dialysate floor with the membrane in touch with the dialysate. Though the machine’s patented design could be very easy, the easy-to-use Slide-A-Lyzer MINI Dialysis Unit is a useful software for functions, like equilibrium aggressive dialysis, for which solely 10-100 µl samples can be found.

Dialysis Price and Pattern Restoration The three.5K Slide-A-Lyzer MINI Dialysis Unit was used for salt discount evaluation. Samples of 5-100 µl of 1 M NaCl had been positioned within the Slide-A-Lyzer MINI Dialysis Unit and dialyzed towards 1 L of water for 10 minutes. To get better the smallest (5 µl and 10 µl) volumes from the Slide-A-Lyzer MINI Dialysis Unit, the machine was tilted and gently tapped on the underside edge to pool the pattern. NaCl requirements and samples had been diluted in 50 ml ultrapure water and conductivity was measured (Cole-Parmer). The Slide-A-Lyzer MINI Dialysis Unit dialyzes effectively (Determine 1). Dialysis price of 100 µl of 5 M NaCl was additionally analyzed by conductivity (Determine 2). In a 3rd experiment, the speed of pH alternate within the Slide-A-Lyzer MINI Dialysis Unit was decided and can also be fast. In lower than 10 minutes, 100 µl of IgG Elution Buffer, pH 2.8 is transformed to pH 9.4 by dialysis towards 1 L of BupH™ Carbonate-Bicarbonate Buffer, pH 9.4 (information not proven). 100% 80% % Residual NaCl

Highlights: • 100% leak-tested Patented design doesn’t allow “wicking” that may happen in do-it-yourself units • Very reasonably priced • Wonderful pattern recoveries The Slide-A-Lyzer MINI Dialysis Unit usually recovers 9-10 µl after dialysis of a ten µl pattern • Time of dialysis drastically diminished Converts 100 µl of pH 2.8 buffer to pH 9.4 dialyzing towards 1 L bicarbonate buffer, pH 9.4 in lower than 10 minutes

60% 40% 20% 0% 5

10

20

30

40

50

60

70

80

90

100

1 M NaCl (µl)

Determine 1. Dialysis effectivity in a Thermo Scientific Slide-A-Lyzer MINI Dialysis Unit. After dialysis towards water for 10 minutes, the residual NaCl is 0% for 5-10 µl samples,

1. Apply pattern with a pipette.

2. Place the Slide-A-Lyzer MINI Dialysis Unit into the float.

% Residual NaCl

100% 75% 50% 25% 0% 0

20

40

60

80

100

Time (Minutes)

Determine 2. Dialysis time course in a Thermo Scientific Slide-A-Lyzer MINI Dialysis Unit. 100 µl of 5 M NaCl was dialyzed for as much as 100 minutes in a Slide-A-Lyzer MINI Dialysis Unit towards 1 L of water. Inside 10 minutes, solely 50% of the NaCI remained; by 100 minutes, no NaCI remained. Ren, J. and Chaires, J.B. (2001). Speedy screening of structurally selective ligand binding to nucleic acids. Methodology Enzymol. 340, 99-108.

3. Insert the float into the beaker containing the dialysate.

4. Get better pattern.

See ordering data on pages 8-10.

4

For extra data, or to obtain a product instruction booklet, go to www.thermo.com/pierce

Thermo Scientific Slide-A-Lyzer Dialysis Cassettes Require simply half the time of dialysis tubing Highlights: • > 95% pattern restoration Pattern quantity stays seen all through dialysis • No knots or clamps to loosen and leak Safe design prevents pattern loss attributable to leaks • Inflexible body permits easy pattern withdrawal Eradicating each final drop is simple – even for scientists who’ve by no means earlier than carried out dialysis • Excessive floor space/pattern quantity ratio will dialyze twice as quick as dialysis through standard tubing Patented Cassette design spreads the pattern over a big floor space and the double membrane promotes quick dialysis

Membrane Outer Plastic Casing Outer Plastic Case Half

Membrane

Silicone-like Gasket

Silicone-like Gasket

Membrane

Inlet Port A silicone-like gasket, which is sort of completely hidden from view after the Cassette has been assembled, prevents the Slide-A-Lyzer Dialysis Cassette from leaking.

Outer Plastic Case Half 1. Take away a Cassette from the protecting pouch. Fill the Cassette cavity along with your pattern by one of many information inlets within the nook of the Cassette. With the syringe nonetheless inserted into the cavity, draw up on the syringe to take away air.

2. Connect a flotation buoy and dialyze. Every buoy serves as an efficient flotation machine and likewise as a handy bench-top stand for the Cassette.

3. Inject the Cassette chamber with air and withdraw your dialyzed pattern from the Cassette.

Determine 1

Determine 2

The Slide-A-Lyzer Dialysis Cassette (exploded view) seems like a sandwich (Determine 1). When the entire items are compressed collectively (Determine 2), the outer plastic case halves are welded collectively sonically, hermetically sealing an inside chamber that may be accessed solely through a syringe needle inserted by the inlet port. As a result of the inert gasket is 10 mm huge, the needle path is sealed utterly and tightly when the syringe is withdrawn.

Quantitative Pattern Restoration Three pattern quantity batches of water (0.5 ml, 1.7 ml and three.0 ml) had been loaded and recovered per the respective producer’s directions in a Slide-A-Lyzer Dialysis Cassette and traditional dialysis tubing to find out the volumes of restoration. Water quantity recovered was decided gravimetrically. The next desk summarizes the outcomes: The Slide-A-Lyzer Dialysis Cassette is the product of alternative for quickly dialyzing pattern volumes from 100 µl to 30 ml. The Cassette’s patented design, which gives a most floor space/pattern quantity ratio, permits for glorious pattern recoveries. Not like customary flat tubing, the modern Cassette doesn’t require the usage of knots or clips that may result in leaking and pattern loss.

Common Pattern Quantity Restoration

Pattern Quantity Loaded

Thermo Scientific Slide-A-Lyzer Dialysis Cassette % Quantity Restoration

Conventional Dialysis Tubing % Quantity Restoration

3.0 ml

99.47

92.32

1.7 ml

99.30

93.12

0.5 ml

98.76

87.51

See ordering data on pages 7-11.

To order, name 800-874-3723 or 815-968-0747. Exterior america, contact your native department workplace or distributor.

5

Thermo Scientific SnakeSkin Dialysis Tubing Keep away from the hassles of large-sample dialysis utilizing flat tubing Tubing Specs Membrane Kind: Regenerated cellulose Glycerol Content material: Varies with MWCO membrane Sulfur Content material: 0.1%-0.15% Heavy Metals Content material: Hint Tubing Nominal Dry Thickness* 3.5K MWCO 1.0 mils 7K MWCO

1.2 mils

10K MWCO

0.9 mils

*1 mil = 25 microns

6

We suggest closure utilizing SnakeSkin Dialysis Tubing Clips (offered individually). To make use of the clips, minimize the specified size of tubing, fold one finish over twice and apply a clip. Add the pattern by the second finish of the tubing, fold over twice and fix the second clip. As an alternative choice to these clips, SnakeSkin Dialysis Tubing will also be closed with knots. Dip two to a few inches of 1 finish of the tubing into water or buffer and tie a knot within the moist membrane. (Dipping is required to guarantee a great seal on the knot level.) Add the pattern to the open, dry finish and tie a knot at this finish. As a result of the pattern shortly hydrates the membrane, there is no such thing as a have to pre-wet the second finish of the tubing. The pleating course of doesn’t change the tubing’s MWCO. Additionally, any low MW contaminants current are eliminated throughout the dialysis course of. As a result of SnakeSkin Dialysis Tubing is created from the identical kind of regenerated cellulose as flat tubing, its dialysis efficiency matches that of standard tubing.

Conventional flat dialysis tubing is troublesome to open and sometimes requires a presoak in water or buffer earlier than it may be used. Dealing with the tubing after the presoak step might be messy and awkward. Thermo Scientific SnakeSkin Dialysis Tubing was developed to simplify largesample dialysis. SnakeSkin Dialysis Tubing is open, regenerated cellulose dialysis tubing that’s pleated (compressed) right into a hole stick. It’s provided in eight-inch sticks containing 35 toes of twenty-two mm inner diameter (I.D.) tubing, equal to 10.5 meters of 34 mm dry flat width tubing. SnakeSkin Dialysis Tubing can be utilized for 15-100 ml samples. The hydrated tubing will maintain ~3.7 ml of pattern per centimeter of size.

SnakeSkin Dialysis Tubing is accessible in three MWCOs: 3.5K, 7K and 10K. The product is saved in its authentic packaging at room temperature, though refrigerated storage may be used. Correctly saved membrane is steady for at the very least one yr.

68011

SnakeSkin Dialysis Tubing Clips

6/pkg.

The pleated format of SnakeSkin Dialysis Tubing makes it straightforward to open and able to use, streamlining dialysis preparation. To make use of it, a researcher merely pulls out the required size of tubing, cuts it off and applies a closure. The pattern is then added by the opposite finish of the dry tubing and the second closure is utilized.

66432

Slide-A-Lyzer Buoys for 12 ml Slide-A-Lyzer Cassettes

10/pkg.

Ordering Data Product # Description

MWCO

Pkg. Dimension

68035

SnakeSkin Dialysis Tubing

3.5K

22 mm dry I.D. x 35 toes*

68700

SnakeSkin Dialysis Tubing

7K

22 mm dry I.D. x 35 toes*

68100

SnakeSkin Dialysis Tubing

10K

22 mm dry I.D. x 35 toes*

*Equal to 10.5 meters of 34 mm dry flat width tubing.

Product Equipment Product # Description

See a demo of SnakeSkin Tubing at www.thermo.com/dialysis

For extra data, or to obtain a product instruction booklet, go to www.thermo.com/pierce

Pkg. Dimension.

2K MWCO Membrane Merchandise Specs

100 % Residual NaCI

Membrane Composition: Regenerated cellulose synthesized by the Viscose technique Hydration Required Earlier than Use: 2 minutes

75 50 25 0

Glycerol Content material: None

0

1

Sulfur Content material: 0.169% Heavy Metals Content material: Hint

2 3 Time in hours

4

5

Desalting price of the membrane for salts. Sodium chloride (1 M) in water was dialyzed at 4°C and the speed of elimination of NaCl was decided by measuring the conductivity of the retentate at completely different time intervals.

Ordering Data

100

2K MWCO – 0.5 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description Capability

Pkg. Dimension

66205

10/pkg.

Slide-A-Lyzer Dialysis Cassette

0.2-0.5 ml

2K MWCO – 3 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description Capability

Pkg. Dimension

66203

10/pkg.

Slide-A-Lyzer Dialysis Cassette

0.5-3 ml

80

% Retention

1355 Da (63%) Vitamin B12 60

1422 Da (79%) Bacitracin

2K MWCO – 12 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description Capability

Pkg. Dimension

1670 Da (85%) T Kinase Peptide

66212

8/pkg.

1897 Da (93%) Biotin-TPK

40

20

Slide-A-Lyzer Dialysis Cassette

3-12 ml

2067 Da (90%) Ce Peptide

2K MWCO – 30 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description Capability

Pkg. Dimension

2531 Da (88%) Insulin Chain A

66230

6/pkg.

0 Mannequin Programs

Characterization of membrane pore dimension. Vitamin B12, bacitracin, tyrosine kinase peptide 1, biotin-TPKs substrate, protein kinase Ce (PKCe) peptide substrate and insulin chain A mannequin methods (0.5-1 mg/ml) in both saline or 0.2 M carbonate bicarbonate buffer pH 9.4 had been dialyzed in a single day (17 hours) at 4°C. The quantity of retentate was estimated utilizing both the Pierce BCA Protein Assay or absorption at 360 nm (for vitamin B12).

Slide-A-Lyzer Dialysis Cassette

12-30 ml

Product Equipment Product # Description

Pkg. Dimension

66430

10/pkg.

Slide-A-Lyzer Buoys Holds one 0.1-0.5 ml or 0.5-3 ml cassette.

66431

Slide-A-Lyzer Carousel Buoy

1/pkg.

Holds ten 0.1-0.5 ml or 0.5-3 ml cassettes.

66432

Slide-A-Lyzer Buoys

8/pkg.

Holds one 3-12 ml cassette.

View or request our FREE Cell Lysis Options Handbook as we speak! The 36-page, full-color handbook gives technical and product data associated to protein extraction, cell fractionation, DNA extraction, detergents and protein refolding.

66494

Slide-A-Lyzer Syringe (1 ml)

10/pkg.

66490

Slide-A-Lyzer Syringe (5 ml)

10/pkg.

66493

Slide-A-Lyzer Syringe (20 ml)

10/pkg.

Every syringe comes with 18-gauge 1-inch beveled needles.

Go browsing to www.thermo.com/pierce or name 800-874-3723 or 815-968-0747 to request your free copy as we speak! Exterior america, contact your native department workplace or distributor.

To order, name 800-874-3723 or 815-968-0747. Exterior america, contact your native department workplace or distributor.

7

3.5K MWCO Membrane Merchandise Specs Membrane Composition: Regenerated cellulose synthesized by the Viscose technique Hydration Required Earlier than Use: 30 seconds

Ordering Data 3.5K MWCO – Slide-A-Lyzer MINI Dialysis Items Product # Description Capability

Pkg. Dimension

69554

10-100 µl

10/pkg.

10-100 µl

10/pkg.

10-100 µl

50/pkg.

10-100 µl

250/pkg.

Enough caps are included.

69558

Glycerol Content material: Hint Sulfur Content material: 0.1%–0.15%

Slide-A-Lyzer MINI Dialysis Unit Plus Microtubes Slide-A-Lyzer MINI Dialysis Items and Float Enough caps are included.

69550

Heavy Metals Content material: Hint

Slide-A-Lyzer MINI Dialysis Unit Enough caps are included.

69552

Slide-A-Lyzer MINI Dialysis Unit Enough caps are included.

3.5K MWCO – 0.5 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description Capability Pkg. Dimension 66333

Slide-A-Lyzer Dialysis Cassette

0.1-0.5 ml

10/pkg.

66335

Slide-A-Lyzer Dialysis Cassette Equipment

0.1-0.5 ml

Equipment

Incorporates 10 cassettes, 10 buoys and 10 syringes. 100

3.5K MWCO – 3 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description Capability

% Retention

80

60

Pkg. Dimension

66330

Slide-A-Lyzer Dialysis Cassette

0.5-3 ml

10/pkg.

66332

Slide-A-Lyzer Dialysis Cassette Equipment

0.5-3 ml

Equipment

Incorporates 10 cassettes, 10 buoys and 10 syringes.

40

Vitamin B12 (1,355 MW) Insulin B Chain (3.5K MW) Insulin (5K MW)

20

0

3.5K MWCO – 12 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description Capability

Pkg. Dimension

66110

Slide-A-Lyzer Dialysis Cassette

3-12 ml

8/pkg.

66107

Slide-A-Lyzer Dialysis Cassette Equipment

3-12 ml

Equipment

3.5K MWCO

Pattern retention by the three.5K MWCO Thermo Scientific Slide-A-Lyzer Dialysis Cassette membrane. Identified MW requirements had been dissolved at a focus of 1 mg/ml in both 0.15 M sodium chloride or 0.2 M carbonate-bicarbonate buffer, pH 9.4 (Product # 28382). Rotating cells had been assembled with the nominal 3.5K MWCO membranes. One half of the cell was full of MW customary resolution and the opposite half was full of an equal quantity of the plain diluent. Cells had been rotated in a single day at 100 rpm.

3.5K MWCO – 30 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description Capability Pkg. Dimension 66130

Slide-A-Lyzer Dialysis Cassette

3.5K MWCO – SnakeSkin Dialysis Tubing Product # Description 68035

SnakeSkin Dialysis Tubing Equal to 10.5 meters of 34 mm dry width.

8

For extra data, or to obtain a product instruction booklet, go to www.thermo.com/pierce

12-30 ml

6/pkg.

Pkg. Dimension 22 mm dry I.D. x 35 ft

7K MWCO Membrane Merchandise Specs Membrane Composition: Regenerated cellulose synthesized by the Viscose technique Hydration Required Earlier than Use: 30 seconds for low-volume samples Glycerol Content material: 13%

Ordering Data 7K MWCO – Slide-A-Lyzer MINI Dialysis Items Product # Description

Capability

Pkg. Dimension

69560

10-100 µl

50/pkg.

10-100 µl

250/pkg

Slide-A-Lyzer MINI Dialysis Unit Enough caps are included.

69562

Slide-A-Lyzer MINI Dialysis Unit Enough caps are included.

Sulfur Content material: 0.1%–0.15%

7K MWCO – 0.5 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description Capability

Pkg. Dimension

Heavy Metals Content material: Hint

66373

Slide-A-Lyzer Dialysis Cassette

0.1-0.5 ml

10/pkg.

66375

Slide-A-Lyzer Dialysis Cassette Equipment

0.1-0.5 ml

Equipment

Incorporates 10 cassettes, 10 buoys and 10 syringes.

7K MWCO – 3 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description Capability

Pkg. Dimension

66370

Slide-A-Lyzer Dialysis Cassette

0.5-3 ml

10/pkg.

66372

Slide-A-Lyzer Dialysis Cassette Equipment

0.5-3 ml

Equipment

Incorporates 10 cassettes, 10 buoys and 10 syringes.

100

80

7K MWCO – 12 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description Capability

Pkg. Dimension

66710

Slide-A-Lyzer Dialysis Cassette

3-12 ml

8/pkg.

66707

Slide-A-Lyzer Dialysis Cassette Equipment

3-12 ml

Equipment

% Retention

Incorporates 8 cassettes, 8 buoys and 10 syringes. 60

7K MWCO – SnakeSkin Dialysis Tubing Product # Description

40

Vitamin B12 (1,355 MW) Insulin B Chain (3.5K MW) Insulin (5K MW) Cytochrome C (12.3K MW)

20

0 7K MWCO

Pattern retention by the 7K MWCO Thermo Scientific Slide-A-Lyzer Dialysis Cassette membrane. Identified MW requirements had been dissolved at a focus of 1 mg/ml in both 0.15 M sodium chloride or 0.2 M carbonate-bicarbonate buffer, pH 9.4 (Product # 28382). Rotating cells had been assembled with the nominal 7K MWCO membranes. One half of the cell was full of MW customary resolution and the opposite half was full of an equal quantity of the plain diluent. Cells had been rotated in a single day at 100 rpm.

68700

SnakeSkin Dialysis Tubing Equal to 10.5 meters of 34 mm dry width.

Pkg. Dimension 22 mm dry I.D. x 35 ft

Product Equipment Product # Description

Pkg. Dimension

69588

4/pkg.

Slide-A-Lyzer MINI Dialysis Unit Float Holds 25 MINI Dialysis Items

66430

Slide-A-Lyzer Buoys

10/pkg.

Every buoy holds one 0.1-0.5 ml or 0.5-3 ml cassette.

66431

Slide-A-Lyzer Carousel Buoy

1/pkg.

Every buoy holds ten 0.1-0.5 ml or 0.5-3 ml cassettes.

66432

Slide-A-Lyzer Buoys

8/pkg.

Every buoy holds one 3-12 ml cassette.

66494

Slide-A-Lyzer Syringe (1 ml capability)

10/pkg.

66490

Slide-A-Lyzer Syringe (5 ml capability)

10/pkg.

66493

Slide-A-Lyzer Syringe (20 ml capability)

10/pkg.

Every syringe comes with 18-gauge 1-inch beveled needles.

68011

SnakeSkin Dialysis Tubing Clips

6/pkg.

To order, name 800-874-3723 or 815-968-0747. Exterior america, contact your native department workplace or distributor.

9

10K MWCO Membrane Merchandise Specs Membrane Composition: Regenerated cellulose synthesized by the Viscose technique

10K MWCO – 0.5 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description Capability

Pkg. Dimension

66383

Slide-A-Lyzer Dialysis Cassette

0.1-0.5 ml

10/pkg.

66384

Slide-A-Lyzer Dialysis Cassette

0.1-0.5 ml

5 x 10/pkg.

66385

Slide-A-Lyzer Dialysis Cassette Equipment

0.1-0.5 ml

Equipment

Hydration Required Earlier than Use: 30 seconds Glycerol Content material: 21%

Incorporates 10 cassettes, 10 buoys and 10 syringes.

Sulfur Content material: 0.05%

10K MWCO – 3 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description Capability

Pkg. Dimension

Heavy Metals Content material: Hint

66380

Slide-A-Lyzer Dialysis Cassette

0.5-3 ml

10/pkg.

66381

Slide-A-Lyzer Dialysis Cassette

0.5-3 ml

5 x 10/pkg.

66382

Slide-A-Lyzer Dialysis Cassette Equipment

0.5-3 ml

Equipment

Incorporates 10 cassettes, 10 buoys and 10 syringes.

10K MWCO – 12 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description Capability

Pkg. Dimension

66810

Slide-A-Lyzer Dialysis Cassette

3-12 ml

8/pkg.

66811

Slide-A-Lyzer Dialysis Cassette

3-12 ml

5 x 10/pkg.

66807

Slide-A-Lyzer Dialysis Cassette Equipment

3-12 ml

Equipment

Incorporates 8 cassettes, 8 buoys and 10 syringes.

100

% Retention

80

60

40 Insulin B Chain (3.5K MW) Insulin (5K MW) Cytochrome C (12,384 MW)

20

10K MWCO – 30 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description Capability

Pkg. Dimension

66830

12-30 ml

6/pkg.

λ Irradiated 10K MWCO Membrane Product # Description

Capability

Pkg. Dimension

66454

Slide-A-Lyzer Dialysis Cassette

0.1-0.5 ml

10/pkg.

66455

Slide-A-Lyzer Dialysis Cassette

0.5-3 ml

10/pkg.

66453

Slide-A-Lyzer Dialysis Cassette

3-12 ml

8/pkg.

66456

Slide-A-Lyzer Dialysis Cassette

12-30 ml

8/pkg.

Slide-A-Lyzer Dialysis Cassette

0 10K MWCO

Pattern retention by the 10K MWCO Thermo Scientific Slide-A-Lyzer Dialysis Cassette membrane. Identified MW requirements had been dissolved at a focus of 1 mg/ml in both 0.15 M sodium chloride or 0.2 M carbonate-bicarbonate buffer, pH 9.4 (Product # 28382). Rotating cells had been assembled with the nominal 10K MWCO membranes. One half of the cell was full of MW customary resolution and the opposite half was full of an equal quantity of the plain diluent. Cells had been rotated in a single day at 100 rpm.

Ordering Data 10K MWCO – Slide-A-Lyzer MINI Dialysis Items Product # Description 69574

Capability

Slide-A-Lyzer MINI Dialysis Unit Plus 10-100 µl Microtubes

Pkg. Dimension

Slide-A-Lyzer MINI Dialysis Unit Slide-A-Lyzer MINI Dialysis Unit Slide-A-Lyzer MINI Dialysis Unit Plus Float

12-30 ml

22 mm dry I.D. x 35 ft

Product Equipment Product # Description

Pkg. Dimension

69588

Slide-A-Lyzer MINI Dialysis Unit Float

4/pkg.

66430

Slide-A-Lyzer Buoys

10/pkg.

Holds one 0.1-0.5 ml or 0.5-3 ml cassette.

10-100 µl

50/pkg.

66432

Slide-A-Lyzer Carousel Buoy

1/pkg.

Slide-A-Lyzer Buoys

8/pkg.

Holds one 3-12 ml cassette. 10-100 µl

250/pkg.

10-100 µl

Equipment/10 items

66494

Slide-A-Lyzer Syringe (1 ml capability)

10/pkg.

66490

Slide-A-Lyzer Syringe (5 ml capability)

10/pkg.

66493

Slide-A-Lyzer Syringe (20 ml capability)

10/pkg.

Every syringe comes with 18-gauge 1-inch beveled needles.

Enough caps are included.

68011

10

Pkg. Dimension

Holds ten 0.1-0.5 ml or 0.5-3 ml cassettes.

Enough caps are included.

69576

SnakeSkin Dialysis Tubing Equal to 10.5 meters of 34 mm dry width.

66431

Enough caps are included.

69572

68100

10/pkg.

Enough caps are included.

69570

10K MWCO – SnakeSkin Dialysis Tubing Product # Description

SnakeSkin Dialysis Tubing Clips

For extra data, or to obtain a product instruction booklet, go to www.thermo.com/pierce

6/pkg.

20K MWCO Membrane Merchandise Specs Membrane Composition: Regenerated cellulose synthesized by the Viscose technique Hydration Required Earlier than Use: 2 minutes Glycerol Content material: None Sulfur Content material: 0.04% Heavy Metals Content material: Hint

View or request our FREE 2D Mass Spec Pattern Prep Technical Handbook as we speak! This 30-page handbook breaks the 2-D/Mass Spec course of into 5 logical steps after which gives protocols and technical and product data to assist maximize outcomes. The handbook gives background, useful hints and troubleshooting recommendation for cell lysis, 2-D pattern prep, detection, mass spec pattern prep and downstream functions. Thrilling new merchandise embrace Zeba Micro Desalt Spin Columns, ProteoSeek Albumin and IgG Elimination Kits, Imperial™ Protein Stain, The In-Answer Tryptic Digest and Guanidination Equipment, and Deuterated (Heavy) Crosslinkers. Go browsing to www.thermo.com/pierce or name 800-874-3723 or 815-968-0747 to request your free copy as we speak! Exterior america, contact your native department workplace or distributor.

Ordering Data 20K MWCO – 0.5 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description Capability Pkg. Dimension

% Protein Retention

100

66005

Slide-A-Lyzer Dialysis Cassette

0.1-0.5 ml

10/pkg.

90

20K MWCO – 3 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description Capability

Pkg. Dimension

80

66003

10/pkg.

70 Insulin B Chain (3.5K MW) Cytochrome C (12.5K MW) Lysozyme (14K MW) Myoglobin (17K MW)

60

50 Mannequin Programs

Characterization of membrane pore dimension. Insulin B chain, cytochrome C, lysozyme and myoglobin had been dialyzed in a single day (17 hours) at 4°C in PBS pH 7.4. The quantity of retentate was estimated utilizing the Pierce BCA Protein Assay.

Slide-A-Lyzer Dialysis Cassette

0.5-3 ml

20K MWCO – 12 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description Capability

Pkg. Dimension

66012

8/pkg.

Slide-A-Lyzer Dialysis Cassette

3-12 ml

20K MWCO – 30 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description Capability

Pkg. Dimension

66030

6/pkg.

Slide-A-Lyzer Dialysis Cassette

12-30 ml

Product Equipment Product # Description

Pkg. Dimension

69588

Slide-A-Lyzer MINI Dialysis Unit Float

4/pkg.

66430

Slide-A-Lyzer Buoys

10/pkg.

Holds one 0.1-0.5 ml or 0.5-3 ml cassette.

66431

Slide-A-Lyzer Carousel Buoy

1/pkg.

Holds ten 0.1-0.5 ml or 0.5-3 ml cassettes.

66432

Slide-A-Lyzer Buoys

8/pkg.

Holds one 3-12 ml cassette.

66494

Slide-A-Lyzer Syringe (1 ml capability)

10/pkg.

66490

Slide-A-Lyzer Syringe (5 ml capability)

10/pkg.

66493

Slide-A-Lyzer Syringe (20 ml capability)

10/pkg.

Every syringe comes with 18-gauge 1-inch beveled needles.

68011

SnakeSkin Dialysis Tubing Clips

6/pkg.

To order, name 800-874-3723 or 815-968-0747. Exterior america, contact your native department workplace or distributor.

11

The RED Gadget for Speedy Equilibrium Dialysis A reworking know-how for plasma protein-binding assays Figuring out the extent to which a molecule binds to plasma proteins is a important part of drug growth as a result of it influences compound dosing, efficacy, clearance price and potential for drug interactions. This willpower is enabled by equilibrium dialysis, an accepted customary technique for dependable estimation of the nonbound drug fraction in plasma. Though it’s the popular technique, equilibrium dialysis is labor-intensive, time-consuming, cost-prohibitive and troublesome to automate. The RED Gadget† for fast equilibrium dialysis was developed in shut affiliation with the pharmaceutical trade to particularly tackle these points, accelerating lead optimization and lowering attrition price. The RED System consists of disposable tube inserts and a 96-well Teflon® Base Plate. The distinctive design of the bottom plate gives compatibility with automated liquid dealing with methods whereas the massive dialysis floor space of the tube inserts accelerates equilibrium. The RED Gadget has been extensively validated for plasmabinding assays producing outcomes according to these reported within the literature (Desk 1). Utilizing the RED Gadget to measure Warfarin binding to plasma (human and rat) proteins at two concentrations of 1 and 10 µM, the RED Gadget produced outcomes with minimal intra-experimental variability (Determine 1). The RED System provides vital enhancements within the ease of use, time necessities, versatility and product reliability in comparison with opponents (Desk 2). The RED Gadget Allows: • Willpower of free vs. certain drug to plasma proteins • Pharmacokinetics research • Formulation of drug dosage for in vivo research • Drug-to-drug interplay research • Choice standards throughout drug lead optimization

12

Highlights: • Ease of use Disposable tubes require no presoaking, meeting or specialised gear • Brief incubation time Equilibrium might be reached in as few as three hours because of the excessive membrane surface-to-volume ratio • 96-well format Appropriate for automated liquid handlers • Versatile Can be utilized for the specified variety of assays (one to 48 assays/ plate) with out losing your entire plate • Strong Compartmentalized design eliminates potential for crosstalk or leakage • Reproducible and correct Validated for plasma-binding assays, producing outcomes according to these reported within the literature (Desk 1) • Versatile The high-grade Teflon Base Plate is chemically inert, eliminating nonspecific binding and danger of contamination • Validated Every lot is functionally examined in a protein-binding assay for assured efficiency • Handy The RED Gadget membrane has a MWCO of 8K; different MWCO membranes can be found upon request Desk 1. Comparability of outcomes obtained utilizing the RED Gadget with values reported within the literature. % Sure Compound

Literature Worth

RED Gadget

Ranitidine1

10-19

17

87-96

84

99

99

99

99

2

Propranolol 3

Warfarin

1

Naproxen

1. Jusko, W.J. and Gretch, M. (1976). Plasma and tissue protein binding of medication in pharmacokinetics. Drug Metab. Rev., 5(1), 42-139. 2. Colangelo, P.M., et al. (1992). Age and propranolol stereoselective disposition in people. Clin. Pharmacol. Ther., 51, 489-94. 3. Chan, E., et al. (1994). Disposition of warfarin enantiomers and metabolites in sufferers throughout a number of dosing with rac-warfarin. Brit. J. Clin. Pharmacol., 36, 563-569.

For extra data, or to obtain a product instruction booklet, go to www.thermo.com/pierce

The RED Gadget for Speedy Equilibrium Dialysis A reworking know-how for plasma protein-binding assays Human plasma protein binding of Warfarin

Rat plasma protein binding of Warfarin 0.01

Drug Fraction Free

Drug Fraction Free

0.01 1 µM 10 µM

0.008 0.006 0.004 0.002

1 µM 10 µM

0.008 0.006 0.004 0.002 0

0 0

1

2

3

4

0

5

1

2

3

4

5

Time (Hour)

Time (Hour)

Determine 1. The RED Gadget binds plasma proteins. Greater than 99% of Warfarin was persistently certain to plasma protein displaying minimal intra-experimental variability. Three replicate RED Gadget inserts had been arrange for every examined time level. Warfarin options at 1 µM or 10 µM had been made within the plasma of alternative and added to the insert pattern chamber. PBS was added to the buffer chamber. At every time level (1, 2, 3 and 4 hours), 50 µl was faraway from the plasma and the buffer chambers and transferred to separate wells of a deep nicely plate.

After on a regular basis factors had been collected, 50 µl of clean plasma was added to each buffer pattern and 50 µl buffer was added to each plasma pattern. After precipitation buffer was added, vortexed and centrifuged, the supernatants had been analyzed by LC/MS/MS (API4000). An ordinary curve of the drug of curiosity was ready together with the samples. The focus of every pattern was decided from the usual curve.

Desk 2. A comparability of important attributes for equilibrium units.†† Hours to succeed in Equilibrium

Leakage

Disposable

Labor Depth

Automation Accessible

Quantity Shift

4

None

Sure

Sure

None

Multi-Equilibrium Dialyzer (Harvard Equipment)

3-4

Minimal

No

••••

No

Minimal

96-well Equilibrium DIALYZER (Harvard Equipment)

16

20%

Sure

•••

Potential

Sure

96-well Micro Equilibrium Dialysis Block (HTDialysis, LLC)

6

Some

No

•••

Potential

Sure

24-Multiwell Dialysis (BD Biosciences)

24

Not measured

Sure

••

Potential

Not measured

Gadget (Supply) RED (Speedy Equilibrium Dialysis) Gadget (Thermo Scientific)

†† Li, S.,1 Xiong, B.,2 Huang, T.,2 Li, L.,2 Donovan, J.,3 Lee, F.,1 Yu, S.,1 Miwa, G.1 and Yang, H.1 Validation of a novel fast equilibrium dialysis (RED) machine for prime throughput plasma protein binding willpower. 1. DMPK/Drug Security & Disposition; 2. Linden Bioscience, 35A Cabot Highway, Woburn, MA 01801, USA; and three. Course of Know-how, Millennium Prescription drugs, Inc., 40 Landsdowne Avenue, Cambridge, MA 02139 USA.

1. Place RED Gadget Inserts into the Teflon Plate.

2. Place 200 µl of pattern (combination of three. Add applicable quantity of buffer compound with plasma at applicable to the buffer chamber. Cowl plate and concentrations) within the plasma chamber incubate. Equilibrium is commonly reached indicated by the pink ring. in 4 hours or much less.

4. After dialysis, take away equal volumes from each chambers. Proceed to pattern preparation earlier than LC/MS/MS evaluation.

Determine 2. Schematic protocol for the RED Gadget.

Ordering Data Product # Description

Pkg. Dimension

89809

RED Gadget Inserts

50/pack

89810

RED Gadget Inserts

250/case

89811

Teflon Base Plate

1 plate

89812

RED Gadget Insert Elimination Instrument

1 remover

† The RED Gadget is manufactured by Linden Bioscience. Patent Pending on RED Gadget by Linden Bioscience.

To order, name 800-874-3723 or 815-968-0747. Exterior america, contact your native department workplace or distributor.

13

Thermo Scientific Slide-A-Lyzer Concentrating Answer Thermo Scientific Slide-A-Lyzer Concentrating Answer is a proprietary, hygroscopic, excessive MW compound that pulls water by dialysis membrane shortly. Different concentrating options focus and contaminate samples with a compound of comparable MW that’s troublesome to take away by dialysis or different means. These contaminants soak up strongly at 280 nm, distorting protein measurements utilizing the tyrosine absorption technique. The Slide-A-Lyzer Concentrating Answer particular formulation is freed from low MW compounds that would cross the membrane to infect the pattern. Many samples will tackle water or buffer throughout the dialysis course of. To return the pattern to its authentic focus, or to pay attention it even additional, the Slide-A-Lyzer Concentrating Answer is good. To pay attention the pattern, the Slide-A-Lyzer Dialysis Cassette containing the pattern is positioned in a small plastic bag containing the concentrating resolution. By diffusion, water and different small molecules are drawn out of the cassette, into the bag. The big molecular dimension of the concentrating resolution prevents it from crossing the membrane and coming into the cassette. Due to this fact, a one-way circulate of water and different small molecules out of the Cassette leads to focus of the pattern.

Highlights: • Dialysis and focus happen in a single machine Avoids protein loss through the use of a single machine • Sooner focus A beginning quantity of three ml is diminished to 0.5 ml in about 75-80 minutes • Simple to make use of Simply pour the Slide-A-Lyzer Concentrating Answer into the small plastic bag offered and drop within the Slide-A-Lyzer Dialysis Cassette containing the pattern • Improved formulation and protocols Improved product makes focus simpler with rockingplatform protocols • The method might be monitored As a result of each the concentrating resolution and the bag are clear, the pattern focus might be simply monitored, one thing that isn’t potential with closed-system centrifuge-type units

Ordering Data Product # Description

Pkg. Dimension

66528

200 ml

Slide-A-Lyzer Concentrating Answer To be used with 0.5-3 ml cassettes.

66529

Slide-A-Lyzer Concentrating Answer

500 ml

To be used with 3-30 ml cassettes.

66530

Slide-A-Lyzer Concentrating Answer

25 ml

To be used with Slide-A-Lyzer MINI Dialysis Items

Quantity Recovered (ml)

3.5 10K 7K 3.5K

3 2.5 2 1.5 1 0.5 0 0

20

40

60

80

100

120

Time (Minutes) The Slide-A-Lyzer Concentrating Answer shortly reduces a beginning quantity of three ml of pattern contained in the Slide-A-Lyzer Dialysis Cassette to 0.5 ml in about 50 minutes. That is corresponding to different focus strategies reminiscent of centrifuge-driven membrane units.

14

Concentrating with the Thermo Scientific Slide-A-Lyzer MINI Dialysis Unit Slide-A-Lyzer Concentrating Answer works on even very small samples utilizing the Slide-A-Lyzer MINI Dialysis Unit. Samples from 10 to 100 µl are positioned within the Slide-A-Lyzer MINI Dialysis Unit after which positioned in a microcentrifuge tube that accommodates Slide-A-Lyzer Concentrating Answer at a minimal ratio of three:1 (Concentrating Answer to pattern).

For extra data, or to obtain a product instruction booklet, go to www.thermo.com/pierce

Thermo Scientific Pierce Microdialyzer System Dialyze a number of small quantity samples with minimal protein loss – in lower than one hour! Highlights: • Big selection of MWCOs accessible, 1K to 50K • Helpful lid to forestall evaporation • Stir bar to extend dialysis effectivity • Distinctive one-way valve system to supply extra environment friendly regulation of buffer alternate • Design permits entry to dialysate chamber and channels “air bubbles” to a vent valve Purposes: • Purifying membrane proteins • Eradicating protein-modification reagents and crosslinkers • Eradicating oligosaccharides from protein options • Dialyzing previous to electrophoresis • Exchanging, accumulating and/or analyzing buffer throughout dialysis • Blotting for immunochemistry Many dialysis strategies are timeconsuming and end in substantial pattern loss – a important concern when coping with small pattern volumes. The Thermo Scientific Microdialyzer System provides fast, easy strategies for pattern desalting or buffer alternate. These items require solely about one hour to finish the dialysis process, they usually decrease the loss or dilution of valuable samples. They just about eradicate the necessity for awkward, time-consuming dialysis procedures. The Microdialyzer System is good for dialyzing small samples. The Microdialyzer System 100 is designed for the simultaneous dialysis of twelve 20-100 µl samples. The Microdialyzer System is supplied with a lid to forestall pattern evaporation and a novel inlet/outlet system for simple buffer alternate.

Ordering Data Product # Description 66315

Microdialyzer System 100 For simultaneous dialysis of 12 x 20 µl-100 µl samples.

Microdialyzer System Equipment Product # Description 66322

Delrin 2 Pattern Effectively Plate For System 100 For autoclaving functions with Microdialyzer System 100.

Pre-Framed Dialysis Membranes Product # Description

MWCO

Pkg. Dimension

66306

Pre-Framed Dialysis Membranes

1K

10/pkg.

66307

Pre-Framed Dialysis Membranes

3K

10/pkg.

66310

Pre-Framed Dialysis Membranes

8K

10/pkg.

66312

Pre-Framed Dialysis Membranes

20K

10/pkg.

66313

Pre-Framed Dialysis Membranes

50K

10/pkg.

The Microdialyzer System provides minimal protein loss and glorious pattern restoration. As well as, Delrin® Effectively Plates can be found for autoclaving functions with the System 100. Precut and pre-framed cellulose acetate membranes of various MW cutoffs are additionally accessible for added comfort.

Earlier than

Throughout

After

Pattern + Contaminants

Pattern + Contaminants

Pattern

Semipermeable Membrane Solvent

Solvent + Contaminants

Solvent + Contaminants

To order, name 800-874-3723 or 815-968-0747. Exterior america, contact your native department workplace or distributor.

15

Pressured Dialysis for Pattern Focus Concentrates 100 ml all the way down to 12 ml in six hours Thermo Scientific SnakeSkin Dialysis Tubing Pipette Tip (want 2) Lower Down

Methodology 1) Lower off and discard the underside of two pipette suggestions (2.5-5 ml) so SnakeSkin Tubing simply suits by the pipette tip. 2) Insert one pipette by the rubber stopper.

Rubber Stopper with Single Gap

3) Lower off the specified size of SnakeSkin Dialysis Tubing (for bigger volumes, the membrane will prolong above the flask). 4) Thread the SnakeSkin Dialysis Tubing (dry) by the rubber stopper containing the pipette tip. 5) Clip or tie a number of knots within the decrease finish of the SnakeSkin Tubing.

Heavy-walled Buchner Flask Vacuum Pump or Home Vacuum

Thermo Scientific SnakeSkin Dialysis Tubing

Dialysate

6) Pour the pattern to be concentrated by the highest of the open finish of the SnakeSkin Tubing. (Earlier than you utterly fill the SnakeSkin Tubing, place the second pipette tip contained in the SnakeSkin Tubing to create a safe seal between the SnakeSkin Dialysis Tubing and the primary pipette tip.) Fill with remaining pattern. 7) Place 3-4 cm of buffer within the flask. NOTE: Many of the SnakeSkin Tubing won’t be uncovered to buffer. 8) Clip or tie the open finish of the SnakeSkin Dialysis Tubing to make sure a closed vacuum system. 9) Join the facet arm to deal with vacuum.

Small samples dialyze a lot quicker than bigger samples as a result of the focus differential is far increased and the migratory diffusion distance is shorter. With a 100 ml dilute pattern, it’s usually prudent to pay attention all the way down to 12 ml with pressured dialysis utilizing SnakeSkin Tubing earlier than dialysis in a Slide-A-Lyzer Dialysis Cassette. The next pressured dialysis SnakeSkin Tubing utility has been tailored from the tactic described in: Doonan, S. (ed.) (1996). Protein Purification Protocols in Strategies in Molecular Biology, 59, 97-101.

10) Focus pattern till desired quantity is reached. Pattern Outcomes 1) A 1 mg/ml resolution of bovine serum albumin was ready in phosphate-buffered saline, pH 7.4. 2) Roughly 30 cm of SnakeSkin Dialysis Tubing was used and assembled as described beforehand. 3) After six hours, the beginning pattern quantity (100 ml) was concentrated to 12 ml with an estimated protein restoration of 65%.

Evaporation for Pattern Focus Water inside a Slide-A-Lyzer Dialysis Cassette will evaporate. The cassette is ideally fitted to pattern focus through evaporation due to the twin membranes and excessive floor space. Place a pattern within the cassette, then withdraw the air inside. Place two buoys on the cassette as proven on the left. Let your pattern evaporate on the bench prime (utilizing a fan to extend airflow throughout the membrane will pace up the method), ensuring to verify each 10 minutes or much less to forestall evaporation to dryness. When concentrating by evaporating the water out of your pattern, the small molecules (buffer salts, lowering brokers, and so on.) can even be concentrated as a result of no diffusion happens.

16

For extra data, or to obtain a product instruction booklet, go to www.thermo.com/pierce

Desalting Columns and Plates Buffer Alternate

Thermo Scientific Zeba Micro Desalt Spin Columns

96-well Desalt Plates

Gel Filtration Gel filtration entails the chromatographic separation of molecules of various dimensions based mostly on their relative talents to penetrate into Protein Desalting an acceptable stationary part. A Spin Columns chromatographic resin, often consisting of very small, uncharged porous particles in an aqueous resolution, is packed right into a column after which used for the separation. Totally different ranges of separation might be achieved based mostly on the pore dimension of the resin. The resin might be chosen to completely exclude proteins or massive molecules, whereas nonetheless together with small solutes. Giant molecules are excluded from the inner pores of the resin and emerge first from the column within the “void quantity.” The smaller molecules are in a position to penetrate the pores, then progress by the column at a slower price. These smaller molecules emerge from the column after the goal pattern. Desalting and buffer alternate are two of essentially the most broadly used functions of gel filtration chromatography.

Desalting Desalting entails the chromatographic separation of macromolecules within the void quantity from smaller molecules that penetrate the gel mattress. Purposes: • Eradicating salts from protein options • Eradicating phenol from nucleic acid preparations • Separating extra crosslinker from conjugate preparations • Eradicating extra derivatizing brokers from modified proteins • Eradicating unreacted dye from fluorescent antibodies • Eradicating free radiolabel from labeled proteins

Buffer alternate is used to position a protein resolution right into a extra applicable buffer previous to functions reminiscent of electrophoresis, ion alternate or affinity chromatography. In each desalting and buffer alternate, the macromolecular parts are recovered in equilibrium with the identical buffer used to equilibrate the column. If water is used for equilibration, the parts will probably be desalted. If one other buffer is used, a buffer alternate will outcome. Thermo Scientific D-Salt™ Columns (web page 19) are ready-to-use gel filtration columns with a novel stop-flow attribute that forestalls the gel from drying. The columns all work utilizing gravity circulate, so there is no such thing as a want for a pump. A polyethylene disc has been inserted each above and under the column resin mattress, in order that when the buffer is utilized, the meniscus will cease on the prime disc. This enables management of the fraction assortment time and fraction dimension, and it prevents pattern loss.

Thermo Scientific Zeba Desalt Spin Columns Highlights: • Distinctive protein restoration • Extensive product providing accommodates your pattern wants • Simple to make use of with no cumbersome column preparation or equilibration • No screening fractions for protein or ready for protein to emerge by gravity circulate • Minimal pattern dilution Though quite a few strategies and resins for desalting can be found, most have many drawbacks, together with vital pattern loss, lengthy processing instances and the necessity to gather a number of fractions. Zeba Desalt Spin Columns present glorious protein restoration with out the restrictions related to different desalting strategies. Zeba Desalt Spin Columns can be found in micro†, 0.5, 2, 5 and 10 ml codecs and permit processing of samples starting from 2 µl to 4 ml (Desk 1). Desk 1. Really useful pattern volumes for Thermo Scientific Zeba Spin Columns. Resin Mattress

Pattern Quantity

75 µl (micro) column

2-12 µl

0.5 ml column

30-130 µl

2 ml column

200-700 µl

5 ml column

600-2,000 µl

10 ml column

1,500-4,000 µl

96-well

20-100 µl

The straightforward-to-use Zeba Spin-Column Format dramatically improves outcomes over customary drip-column methodologies, eliminating the necessity to watch for samples to emerge by gravity circulate and the necessity to monitor fractions for protein restoration. Zeba Desalt Columns require no chromatographic system, cumbersome column preparation or equilibration they usually can course of a number of samples in ~8 minutes.

To order, name 800-874-3723 or 815-968-0747. Exterior america, contact your native department workplace or distributor.

17

Desalting Columns and Plates Zeba Desalt Spin Columns comprise a high-performance desalting resin that gives distinctive desalting and protein-recovery traits in comparison with different commercially accessible resins (Determine 1). Samples containing as little as 25 µg/ml of protein might be processed, offering distinctive protein restoration and ≥ 95% retention of salts and different small molecules (

Comp. G

Management

Comp. B

Pattern Dimension 200 µl + 50 µl stacker Comp. G

Comp. B

Zeba Column

Management

Pattern Dimension 700 µl

Zeba Column

A.

Pattern 250 µg/ml BSA (66 kDa)

5.2

97%

78%

79%

0.19

0.45

0.73

5.2

90%

56%

29%

Restoration

0.14

0.07

0.14

Conductivity (mho)

B.

Determine 1. Elevated protein restoration with Thermo Scientific Zeba Desalt Spin Columns. Samples of bovine serum albumin (BSA) at Determine 1A. 250 µg/ml and Determine 1B. 25 µg/ml in 1 M NaCl had been desalted with the two ml Zeba Desalt Spin Columns and different industrial desalting resins utilizing comparable codecs. A portion of the recovered pattern (10 µl) was analyzed by SDS-PAGE. The remaining pattern was used for conductivity measurements and Pierce BCA Protein Assay (Product # 23225) was carried out to find out protein focus. Zeba Desalt Resin gives considerably larger protein restoration below all circumstances examined. Conductivity and protein restoration values after desalting are indicated for 250 µg/ml samples.

Thermo Scientific Zeba 96-Effectively Desalt Spin Plates Highlights: • Desalt protein in a single fraction with no dilution • Distinctive protein restoration • Simple to make use of with no cumbersome plate preparation or equilibration • Minimal pattern dilution The brand new Thermo Scientific Zeba 96-Effectively Desalt Spin Plates present high-throughput elimination of salt and small molecules from samples, making ready them for downstream evaluation, together with mass spectrometry, HPLC, capillary electrophoresis, metabolite screening and assay growth. The Zeba 96-Effectively Desalt Spin Plates comprise a high-performance resin that gives distinctive desalting and protein restoration traits. Course of small (20-100 µl) pattern volumes and obtain distinctive protein restoration (Desk 1) and > 95% elimination of salts and different small molecules (

Ordering Data Product # Description

Pkg. Dimension

89877

Zeba Micro Desalt Spin Columns†

25/pack

89878

Zeba Micro Desalt Spin Columns†

50/pack

89882

Zeba Desalt Spin Columns, 0.5 ml

25/pack

89883

Zeba Desalt Spin Columns, 0.5 ml

50/pack

89889

Zeba Desalt Spin Columns, 2 ml

5/pack

89890

Zeba Desalt Spin Columns, 2 ml

25/pack

89891

Zeba Desalt Spin Columns, 5 ml

5/pack

89892

Zeba Desalt Spin Columns, 5 ml

25/pack

89893

Zeba Desalt Spin Columns, 10 ml

5/pack

89894

Zeba Desalt Spin Columns, 10 ml

25/pack

1. Take away the underside seal and stack the desalt plate on prime of a wash plate, then take away the highest seal.

3. Stack the desalt plate on prime of a pattern assortment plate. Apply pattern.

2. Centrifuge for two minutes at 1,000 x g to take away the storage buffer.

4. Centrifuge for two minutes at 1,000 x g.

5. Get better the desalted samples.

Determine 1. Thermo Scientific Zeba 96-Effectively Desalt Spin Plates are straightforward to make use of.

Handee™ Spin Columns (No Resin) Product # Description

Pkg. Dimension

89879

Handee Micro Spin Columns

50/pack

89868

Handee Spin Columns, 0.5 ml

25/pack

BSA (66 kDa)

98.5

> 95

89896

Handee Spin Columns, 2 ml

25/pack

91.5

> 95

89897

Handee Spin Columns, 5 ml

25/pack

α-Lactalbumin (14.1 kDa)

89898

Handee Spin Columns, 10 ml

25/pack

Ubiquitin (8.6 kDa)

85

> 95

Desk 1. Protein restoration and desalting effectivity. Protein

% Protein Restoration

% NaCl Eliminated

Protein samples (1 mg/ml) had been ready in 1.0 M NaCl and 100 µl samples had been desalted utilizing Zeba Desalt Spin Plates. Outcomes had been analyzed by Pierce BCA Protein Assay and conductivity measurements.

18

For extra data, or to obtain a product instruction booklet, go to www.thermo.com/pierce

Desalting Columns and Plates Ordering Data Product # Description

Pkg. Dimension

89807

2 plates

Zeba 96-well Desalt Spin Plates Every nicely accommodates ~550 µl resin slurry and may course of 20-100 µl samples. The package deal accommodates two wash plates and two assortment plates.

89808

Zeba 96-well Desalt Spin Plates

4 plates

Every nicely accommodates ~550 µl resin slurry and may course of 20-100 µl samples. The package deal accommodates two wash plates and 4 assortment plates.

89934

Pierce Desalting Chromatography Cartridges

5 x 1 ml

Every cartridge is full of 1 ml Zeba Desalting Resin. Really useful for processing compounds > 7K MW.

89935

Pierce Desalting Chromatography Cartridges

5 x 5 ml

Every cartridge is full of 5 ml Zeba Desalting Resin. Really useful for processing compounds > 7K MW.

Thermo Scientific D-Salt Columns D-Salt Excellulose™ Desalting Columns Desalt proteins with glorious pattern recoveries.

Thermo Scientific Pierce Desalting Chromatography Cartridges

D-Salt Excellulose Desalting Columns are pre-packed with small, porous cellulose beads. These columns have a moist bead diameter of 100-200 µm and an exclusion restrict of 5 kDa. Excellulose Resin is extraordinarily steady to warmth, freeze-thaw cycles, natural solvents and acidic or primary circumstances. It additionally demonstrates glorious mechanical stability and provides glorious pattern recoveries because of the gradual circulate price.

D-Salt Dextran Desalting Columns Reveals glorious stability and circulate properties. D-Salt Dextran Desalting Columns (5K MWCO) are pre-packed with crosslinked dextran. The moist bead diameter is 50-150 µm. The resin has good rigidity for simple dealing with and glorious circulate properties. The D-Salt Dextran Gel is steady in water, salt options, natural solvents and alkaline or weakly acidic options. It’s heat-stable and might be autoclaved dry or in resolution at a impartial pH for half-hour at 120°C with out affecting its chromatographic properties.

D-Salt Polyacrylamide Desalting Columns Remove the potential for enzymatic degradation and microbial progress. D-Salt Polyacrylamide Desalting Columns are prepacked with porous polyacrylamide beads. D-Salt 6K Columns have a moist bead diameter of 90-180 µm. D-Salt Polyacrylamide 1.8K Desalting Columns have a moist bead diameter of 45-90 µm. These columns are perfect for separating low MW compounds with a fractionation vary of 100-1,800. The D-Salt Polyacrylamide Beads are usually not topic to enzymatic degradation and won’t function a nutrient for microbial progress. The resin could be very hydrophilic, so there must be little interplay with pattern molecules. Contamination with low MW sugars (which can happen with crosslinked dextran) shouldn’t be a priority when utilizing D-Salt Polyacrylamide Resin. Oxidizing brokers might be eliminated with out destroying the assist. This resin is prone to hydrolysis of amide teams below excessive pH circumstances, so an working pH of 2-10 is beneficial at room temperature. The D-Salt Polyacrylamide Resin will also be autoclaved at pH 5.5-6.5 for half-hour at 120°C.

Ordering Data D-Salt Excellulose Desalting Columns Product # Description

MWCO

Pkg. Dimension

20439

D-Salt Excellulose Desalting Columns

5K

5 x 2 ml

20449

D-Salt Excellulose Desalting Columns

5K

5 x 5 ml

D-Salt Dextran Desalting Columns Product # Description

MWCO

Pkg. Dimension

43230

D-Salt Dextran Desalting Columns

5K

5 x 5 ml

43233

D-Salt Dextran Desalting Columns

5K

5 x 10 ml

D-Salt Polyacrylamide Desalting Columns Product # Description

MWCO

Pkg. Dimension

43426

D-Salt Polyacrylamide Desalting Columns

1.8K

5 x 5 ml

43240

D-Salt Polyacrylamide Desalting Columns

6K

5 x 5 ml

43243

D-Salt Polyacrylamide Desalting Columns

6K

5 x 10 ml

89849

Protein Desalting Spin Columns

7K

25 x 0.5 ml

89862

Protein Desalting Spin Columns

7K

50 x 0.5 ml

To order, name 800-874-3723 or 815-968-0747. Exterior america, contact your native department workplace or distributor.

19

Cell Lysis Reagents Desk 1. Thermo Scientific Pierce Cell Lysis Reagents choice information. Dialyze1 Compatibility

Description

Organisms/Samples

B-PER Reagent† 78243, 165 ml 78248, 500 ml

Gram(-) micro organism, S. aureus, H. pylori, E. coli strains BL21(D3)> JM109> DH5a >M15, Archaebacteria, nematodes and Acinetobacter sp.

Sure

Reporter assays, IPs2, Western blot, GST- and histidine-tag purification

B-PER II Reagent 78260, 250 ml (A 2X model of B-PER Reagent)

Gram(-) micro organism, S. aureus, H. pylori, E. coli strains BL21(D3)> JM109> DH5α>M15, Archaebacteria, nematodes and Acinetobacter sp.

Sure

Reporter assays, IPs2, Western blot, GST- and histidine-tag purification

B-PER PBS Reagent 78266, 500 ml

Gram(-) micro organism, S. aureus, H. pylori, E. coli strains BL21(D3)> JM109> DH5α>M15, Archaebacteria, nematodes and Acinetobacter sp.

Sure

Reporter assays, IPs2, Western blot, GST- and histidine-tag purification

Y-PER Reagent 78991, 200 ml 78990, 500 ml

S. cerevisiae, Schizo-saccharomyces pombe, C. albicans, B. subtilis, E. coli, P. pastoris, Strep. avidinii and Acinetobacter sp.

No

IPs2, Western blot, β-Gal enzyme assays, IEF after dialysis, GST- and histidine-tag purification

Y-PER Plus Reagent 78998, 25 ml 78999, 500 ml

Yeast (S. cerevisiae) and Acinetobacter sp.

Sure

GST- and histidine-tag purification, Western blot

M-PER Reagent 78503, 25 ml 78501, 250 ml 78505, 1 L

Cultured mammalian cells, COS-7, NIH3T3, Hepa 1-6, 293, CHO, MDA, MB231 and FM2

Sure

Luciferase, β-Gal (low sign), CAT, kinase assays, ELISAs, immobilized glutathione, Western blot

P-PER Plant Protein Extraction Reagent 89803, Equipment

A number of plant organs (leaf, stem, root, seed and flowers); a number of plant species (Arabidopsis, tobacco, maize, soybeans, peas, spinach, rice and different plant tissues); and recent, frozen and dehydrated plant tissues

No

1-D and 2-D gel electrophoresis, Western blotting, exercise assays and protein affinity purifications*

T-PER Reagent 78510, 500 ml

Coronary heart, liver, kidney and mind

Sure

Luciferase, β-Gal, CAT, kinase assays, Western blot, ELISAs, immobilized glutathione

I-PER Reagent 89802, 250 ml

Baculovirus-infected insect cells grown in suspension or monolayer tradition

No

Western blot, 6xHis-tagged protein purification, protein assays and ion-exchange chromatography

NE-PER Reagent 78833

Tissue: calf liver. Cultured cells: epithelial (HeLa), fibroid (COS-7), kidney (NIH3T3), liver (Hepa 1) and mind (C6)

Mem-PER Reagent 89826

Cultured cells: mind (C6), epithelial (HeLa), fibroblasts (NIH3T3) and yeast (S. cerevisiae)

Yes4

Western blot and 2-D6

Mitochondria Isolation Equipment for Cultured Cells† 89874 Mitochondria Isolation Equipment for Tissue† 89801

Mammalian cells

Yes7

Western blot, 2-D Western blots, electrophoresis. Purposes embrace apoptosis, sign transduction and metabolic research.

RadioImmuno Precipitation Assay (RIPA) Buffer 89900, 100 ml 89901, 250 ml

Cultured mammalian cells and cytoplasmic, membrane and nuclear proteins

Sure

Reporter assays, protein assays, immunoassays and protein purification

Lysosome Enrichment Equipment for Tissues and Cultured Cells 89839

Tissues and Cultured Cells

N/A

2D/MS, electron microscopy, illness profiling, gene expression, sign transduction, and interplay or localization research

Peroxisome Enrichment Equipment for Tissue 89840

Coronary heart, liver, kidney and mind

N/A

2D/MS, electron microscopy, illness profiling, gene expression, sign transduction, and interplay or localization research

Nuclei Enrichment Equipment for Tissue 89841

Coronary heart, liver, kidney and mind

N/A

2D/MS, electron microscopy, illness profiling, gene expression, sign transduction, and interplay or localization research

No (CER) EMSA (if utilizing

Coronary heart, liver, kidney and mind

1. The detergent might be eliminated by dialysis 2. Immunoprecipitation 3. Halt Protease Inhibitor Cocktail, Product # 78410 and 78415 (EDTA-free) 4. Samples ready in Mem-PER Reagent might be dialyzed if the buffer accommodates detergent (e.g., CHAPS), in any other case use PAGEprep ® Advance Equipment (Product # 89888) 5. Slide-A-Lyzer MINI Dialysis Items 6. 2-D Pattern Prep for Nuclear Proteins (Product # 89863) and 2-D Pattern Prep for Membrane Proteins (Product # 89864) are designed utilizing our widespread NE-PER and Mem-PER Reagents. 7. Have to lyse mitochondria first. *Though package works with out liquid nitrogen/freeze-grinding, Dounce homogenization, blade-shearing or glass-bead agitation for cell disruption, it’s appropriate with these different mechanical aids

20

For extra data, or to obtain a product instruction booklet, go to www.thermo.com/pierce

Cell Lysis Reagents Protein Assay Compatibility

Notes

Pierce BCA Assay and Coomassie Plus Assay

Protease inhibitors3 could also be added to forestall protein degradation. Salts, chelating brokers and lowering brokers might be added for extra environment friendly lysis. Don’t exceed 0.5 M NaCl. Higher lysis if cells are frozen in B-PER Reagent.

Pierce BCA Assay and Coomassie Plus Assay after Compat-Ready Protein Assay Reagent Set (Product # 23215) or dilute two to 4 instances

Protease inhibitors3 could also be added to forestall protein degradation. Salts, chelating brokers and lowering brokers might be added for extra environment friendly lysis. Higher lysis if cells are frozen in B-PER Reagent.

Pierce BCA Assay and Coomassie Plus Assay after Compat-Ready Protein Assay Reagent Set (Product # 23215) or dilute two to 4 instances

Protease inhibitors3 could also be added to forestall protein degradation. Salts, chelating brokers and lowering brokers might be added for extra environment friendly lysis. Higher lysis if cells are frozen in B-PER Reagent.

Pierce BCA Assay

Protease inhibitors3 could also be added to forestall protein degradation. Use at room temperature. Double incubation time to be used at 4°C. Use log-phase cells. For stationary part cells, add 0.1 M DTT or 20-50 mM TCEP. Will work with 1 mM EDTA. Doesn’t lyse spores. Can’t use with ion alternate columns.

Pierce BCA Assay and Coomassie Plus Assay

Protease inhibitors3 could also be added to forestall protein degradation. The addition of as much as 2 M NaCl could end in elevated effectivity of lysis and protein yield.

Pierce BCA Assay and Coomassie Plus Assay

Protease inhibitors3 could also be added to forestall protein degradation. Including 150 mM NaCl leads to elevated effectivity of lysis and better protein yield in some cells traces. A PBS rinse of cells previous to lysis removes contaminants reminiscent of phenol pink and will increase protein yield.

Pierce BCA Assay, Lowering Agent-Appropriate Not appropriate with Bradford, Coomassie or Pierce BCA Assay

Equipment lyses most plant cells with out harsh mechanical lysis aids; extraordinarily fibrous tissues reminiscent of woody stems could require mechanical grinding by units not included on this package. P-PER Extracts might be quantified utilizing the Pierce BCA Protein Assay Equipment, Lowering Agent Appropriate (Product # 23250).

Pierce BCA Assay (dilute 1:1) and Coomassie Plus Assay

Protease inhibitors3 could also be added to forestall protein degradation. Mechanical disruption of the tissue remains to be required. May also be used for aesthetic cells.

Pierce BCA Assay

Protease inhibitors3 could also be added to forestall protein degradation.

Pierce BCA Assay and Coomassie Plus Assay (dilute CER Reagent combination 4 instances)

Protease inhibitors3 could also be added to forestall protein degradation. Packed cell vol.: 2 x 106 HeLa cells = 10 µl = 20 mg. Tissue yield (calf liver): 3-4 mg cytoplasmic protein/100 mg tissue; 1-1.5 mg nuclear protein/100 mg tissue. Cell yield (HeLa): 300-400 µg cytoplasmic protein/106 cells; 40-60 µg nuclear protein/106 cells. Constructive controls examined: cytoplasmic (β-Gal, PKC, Hsp90); nuclear (Oct-1, p53, DNA polymerase).

Pierce BCA Assay and Coomassie Plus Assay; hydrophobic part must be dialyzed first; see instruction ebook

Protease inhibitors3 could also be added to forestall protein degradation. Can dialyze towards one other detergent (e.g., CHAPS). Extraction effectivity is mostly > 50% with the cell traces examined (having proteins with as much as two transmembrane segments).

Pierce BCA Assay (after lysis)

Protease inhibitors could also be added to forestall protein degradation. Douncing will improve isolation effectivity vs. detergent alone; nonetheless, a number of samples might be processed concurrently utilizing the reagent-based strategies.

Pierce BCA Assay

Protease inhibitors3 could also be added to forestall proteolysis and preserve phosphorylation of proteins

Coomassie Plus – The Higher Bradford Assay Equipment

Protease inhibitors3 could also be added to forestall proteolysis and preserve phosphorylation of proteins

Coomassie Plus – The Higher Bradford Assay Equipment

Protease inhibitors3 could also be added to forestall proteolysis and preserve phosphorylation of proteins

Coomassie Plus – The Higher Bradford Assay Equipment

Protease inhibitors3 could also be added to forestall proteolysis and preserve phosphorylation of proteins

To order, name 800-874-3723 or 815-968-0747. Exterior america, contact your native department workplace or distributor.

21

Ordering Data Desalting Columns, Plates and Cartridges SnakeSkin Dialysis Tubing

Zeba Spin Columns and Plates U.S. Worth $118

Product # Description 68035 SnakeSkin Dialysis Tubing

MWCO 3.5K

Pkg. Dimension 22 mm dry I.D. x 35 toes*

68700

SnakeSkin Dialysis Tubing

7K

22 mm dry $118 I.D. x 35 toes*

68100

SnakeSkin Dialysis Tubing

10K

22 mm dry $118 I.D. x 35 toes*

*Equal to 10.5 meters of 34 mm dry flat width tubing.

SnakeSkin Dialysis Tubing Accent Product # Description 68011 SnakeSkin Dialysis Tubing Clips

U.S. Pkg. Dimension Worth 6/pkg. $ 47

Product # 89877 89878 89882 89883 89889 89890 89891 89892 89893 89894 89807 89808

Cell Lysis Reagents

Description Zeba Micro Desalt Spin Columns Zeba Micro Desalt Spin Columns Zeba Desalt Spin Columns, 0.5 ml Zeba Desalt Spin Columns, 0.5 ml Zeba Desalt Spin Columns, 2 ml Zeba Desalt Spin Columns, 2 ml Zeba Desalt Spin Columns, 5 ml Zeba Desalt Spin Columns, 5 ml Zeba Desalt Spin Columns, 10 ml Zeba Desalt Spin Columns, 10 ml Zeba 96-well Desalt Plate Zeba 96-well Desalt Plate

U.S. Pkg. Dimension Worth 25/pack $103 50/pack $185 25/pack $ 93 50/pack $169 5/pack $ 36 25/pack $169 5/pack $ 47 25/pack $215 5/pack $ 56 25/pack $255 2 plates $190 4 plates $360

Slide-A-Lyzer Dialysis Cassette Equipment U.S. Pkg. Dimension Worth 4/pkg. $ 20

Product # Description 69588 Slide-A-Lyzer MINI Dialysis Unit Float 66430

Slide-A-Lyzer Buoys

10/pkg.

$ 26

66431

Slide-A-Lyzer Carousel Buoy

1/pkg.

$ 37

66432

Slide-A-Lyzer Buoys

8/pkg.

$ 21

66494

Slide-A-Lyzer Syringe (1 ml)

10/pkg.

$ 21

66490

Slide-A-Lyzer Syringe (5 ml)

10/pkg.

$ 21

66493

Slide-A-Lyzer Syringe (20 ml)

10/pkg.

$ 22

Speedy Equilibrium Dialysis Product # Description 89809 RED Gadget Inserts

U.S. Pkg. Dimension Worth 50/pack $ 350

89810

RED Gadget Inserts

250/case $1,500

89811

Teflon Base Plate

1 plate

89812

RED Gadget Insert Elimination Instrument

1 remover $ 80

$ 320

Slide-A-Lyzer Concentrating Answer U.S. Pkg. Dimension Worth 200 ml $ 80

Product # Description 66528 Slide-A-Lyzer Concentrating Answer

Desalting Chromatography Cartridges Product # Description 89934 Pierce Desalting Chromatography Cartridges 89935 Pierce Desalting Chromatography Cartridges

U.S. Pkg. Dimension Worth 5 x 1 ml $138 5 x 5 ml

$158

Product # 89879 89868 89896 89897 89898

Description Handee Micro Spin Columns Handee Spin Columns, 0.8 ml Handee Spin Columns, 2 ml Handee Spin Columns, 5 ml Handee Spin Columns, 10 ml

Pkg. Dimension 50/pack 25/pack 25/pack 25/pack 25/pack

U.S. Worth $ 49 $ 54 $ 35 $ 40 $ 45

Product # Description 89849 Protein Desalting Spin Columns

U.S. Pkg. Dimension Worth 25/pkg. $ 99

89862

50/pkg.

Protein Desalting Spin Columns

MWCO 5K

U.S. Pkg. Dimension Worth 5 x 2 ml $ 96

66530

Slide-A-Lyzer Concentrating Answer

25 ml

$ 30

20449

D-Salt Excellulose Desalting Columns

5K

5 x 5 ml

$106

43230

D-Salt Dextran Desalting Columns

5K

5 x 5 ml

$ 74

43233

D-Salt Dextran Desalting Columns

5K

5 x 10 ml

$ 81

43426

D-Salt Polyacrylamide Desalting Columns

1.8K

5 x 5 ml

$ 75

43240

D-Salt Polyacrylamide Desalting Columns

6K

5 x 5 ml

$ 71

43243

D-Salt Polyacrylamide Desalting Columns

6K

5 x 10 ml

$ 85

MWCO 9K

U.S. Pkg. Dimension Worth 25/pkg. $129

9K

25/pkg.

$232

20K

25/pkg.

$129

20K

25/pkg.

$232

Microdialyzer System Accent

Pre-Framed Dialysis Membranes

Y-PER Reagent Y-PER Plus Reagent Y-PER Plus Reagent M-PER Reagent M-PER Reagent M-PER Reagent P-PER Plant Protein Extraction Reagent T-PER Reagent I-PER Reagent NE-PER Reagent Mem-PER Reagent Mitochondria Isolation Equipment for Cultured Cells Mitochondria Isolation Equipment for Tissue

500 ml 25 ml 500 ml 25 ml 250 ml 1L Equipment

$249 $ 29 $246 $ 53 $217 $589 $225

500 ml 250 ml Equipment Equipment Equipment

$242 $165 $185 $185 $185

Equipment

$199

RadioImmuno Precipitation Assay (RIPA) Buffer RadioImmuno Precipitation Assay (RIPA) Buffer Lysosome Enrichment Equipment for Tissues and Cultured Cells Peroxisome Enrichment Equipment for Tissue Nuclei Enrichment Equipment for Tissue

100 ml

$ 60

250 ml

$130

Equipment

$250

Equipment

$250

Equipment

$250

78510 89802 78833 89826 89874

89900 89901 89839 89840

D-Salt Desalting Columns Product # Description 20439 D-Salt Excellulose Desalting Columns

U.S. Worth $ 65

78990 78998 78999 78503 78501 78505 89803

$163

$186

Product # Description 66322 Delrin 2 Pattern Effectively Plate For System 100

U.S. Worth $129 $245 $245 $245 $116

89841

Protein Desalting Spin Columns

500 ml

Product # Description 66315 Microdialyzer System 100

Pkg. Dimension 165 ml 500 ml 250 ml 500 ml 200 ml

89801

Slide-A-Lyzer Concentrating Answer

U.S. Worth $499

Description B-PER Reagent B-PER Reagent B-PER II Reagent B-PER PBS Reagent Y-PER Reagent

Handee Spin Columns

66529

Microdialyzer System

Product # 78243 78248 78260 78266 78991

Pierce Protein Concentrators

Product # Description 66306 Pre-Framed Dialysis Membranes

MWCO 1K

U.S. Pkg. Dimension Worth 10/pkg. $119

66307

Pre-Framed Dialysis Membranes

3K

10/pkg.

$119

66310

Pre-Framed Dialysis Membranes

8K

10/pkg.

$119

66312

Pre-Framed Dialysis Membranes

20K

10/pkg.

$119

66313

Pre-Framed Dialysis Membranes

50K

10/pkg.

$119

Product # Description 89884 Pierce Concentrators, 7 ml 89885 Pierce Concentrators, 20 ml 89886 Pierce Concentrators, 7 ml 89887 Pierce Concentrators, 20 ml

Costs listed herein are correct at time of printing. For up-to-date costs , go to www.thermo.com/pierce

Ordering Data 2K MWCO Membrane Merchandise

7K MWCO Membrane Merchandise

0.5 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description 66205 Slide-A-Lyzer Dialysis Cassette

Slide-A-Lyzer MINI Dialysis Items

U.S. Pkg. Dimension Worth 10/pkg. $ 73

3 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description 66203 Slide-A-Lyzer Dialysis Cassette

69562

U.S. Pkg. Dimension Worth 10/pkg. $ 73

12 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description 66212 Slide-A-Lyzer Dialysis Cassette

Product # Description 69560 Slide-A-Lyzer MINI Dialysis Unit

U.S. Pkg. Dimension Worth 8/pkg. $ 94

U.S. Pkg. Dimension Worth 6/pkg. $ 75

3.5K MWCO Membrane Merchandise

Product # Description 66373 Slide-A-Lyzer Dialysis Cassette 66375

U.S. Product # Description Capability Pkg. Dimension Worth 69554 Slide-A-Lyzer MINI 10-100 µl 10/pkg. $ 49 Dialysis Unit Plus Microtubes

Slide-A-Lyzer Dialysis Cassette Equipment

Product # Description 66370 Slide-A-Lyzer Dialysis Cassette Slide-A-Lyzer Dialysis Cassette Equipment

69558

Slide-A-Lyzer MINI 10-200 µl 10/pkg. Dialysis Items and Float

$ 53

69550

Slide-A-Lyzer MINI Dialysis Unit

10-100 µl 50/pkg.

$126

66707

69552

Slide-A-Lyzer MINI Dialysis Unit

10-100 µl 250/pkg.

$384

Slide-A-Lyzer Dialysis Cassette Equipment

66332

Slide-A-Lyzer Dialysis Cassette Equipment

66107

Slide-A-Lyzer Dialysis Cassette Equipment

$102

Slide-A-Lyzer Dialysis Cassette Equipment

U.S. Capability Pkg. Dimension Worth 3-12 ml 8/pkg. $ 94 Equipment

$115

69570

Slide-A-Lyzer MINI Dialysis Unit

10-100 µl 50/pkg.

$126

69572

Slide-A-Lyzer MINI Dialysis Unit

10-100 µl 250/pkg.

$394

69576

Slide-A-Lyzer MINI Dialysis Unit Plus Float

10-100 µl Equipment/10 pkg. $ 53

$ 88

U.S. Capability Pkg. Dimension Worth 0.5-3 ml 10/pkg. $ 73 Equipment

$102

U.S. Capability Pkg. Dimension Worth 3-12 ml 8/pkg. $ 94 3-12 ml

Equipment

Slide-A-Lyzer Dialysis Cassette

3-12 ml

5×10/pkg. $440

66807

Slide-A-Lyzer Dialysis Cassette Equipment

3-12 ml

Equipment

$115

30 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description 66830 Slide-A-Lyzer Dialysis Cassette

U.S. Capability Pkg. Dimension Worth 12-30 ml 6/pkg. $ 76

Product # Description 66454 Slide-A-Lyzer Dialysis Cassette

U.S. Capability Pkg. Dimension Worth 0.1-0.5 ml 10/pkg. $ 99

66455

Slide-A-Lyzer Dialysis Cassette

0.5-3 ml

10/pkg.

$109

66453

Slide-A-Lyzer Dialysis Cassette

3-12 ml

8/pkg.

$100

66456

Slide-A-Lyzer Dialysis Cassette

12-30 ml

6/pkg.

$ 78

0.5 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description 66005 Slide-A-Lyzer Dialysis Cassette

0.1-0.5 ml Equipment

0.5-3 ml

66811

20K MWCO Membrane Merchandise 3-12 ml

U.S. Product # Description Capability Pkg. Dimension Worth 69574 Slide-A-Lyzer MINI 10-100 µl 10/pkg. $ 50 Dialysis Unit Plus Microtubes

$115

30 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description 66130 Slide-A-Lyzer Dialysis Cassette

Equipment

U.S. Capability Pkg. Dimension Worth 3-12 ml 8/pkg. $ 94

λ-Irradiated 10K MWCO Membrane

U.S. Capability Pkg. Dimension Worth 0.5-3 ml 10/pkg. $ 73 0.5-3 ml

Product # Description 66810 Slide-A-Lyzer Dialysis Cassette

U.S. Pkg. Dimension Worth 10/pkg. $ 70

Slide-A-Lyzer MINI Dialysis Items

12 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description 66110 Slide-A-Lyzer Dialysis Cassette

$ 88

U.S. Capability Pkg. Dimension Worth 0.1-0.5 ml 10/pkg. $ 73

3 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description 66330 Slide-A-Lyzer Dialysis Cassette

0.1-0.5 ml Equipment

10K MWCO Membrane Merchandise

0.5 ml Capability Slide-A-Lyzer Dialysis Cassettes

66335

U.S. Capability Pkg. Dimension Worth 0.1-0.5 ml 10/pkg. $ 73

12 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description 66710 Slide-A-Lyzer Dialysis Cassette

Product # Description 66333 Slide-A-Lyzer Dialysis Cassette

$394

3 ml Capability Slide-A-Lyzer Dialysis Cassettes

66372

Slide-A-Lyzer MINI Dialysis Items

10-100 µl 250/pkg.

0.5 ml Capability Slide-A-Lyzer Dialysis Cassettes

30 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description 66230 Slide-A-Lyzer Dialysis Cassette

Slide-A-Lyzer MINI Dialysis Unit

12 ml Capability Slide-A-Lyzer Dialysis Cassettes

U.S. Capability Pkg. Dimension Worth 10-100 µl 50/pkg. $126

U.S. Capability Pkg. Dimension Worth 12-30 ml 6/pkg. $ 76

U.S. Capability Pkg. Dimension Worth 0.1-0.5 ml 10/pkg. $ 73

66384

Slide-A-Lyzer Dialysis Cassette

0.1-0.5 ml 5×10/pkg. $340

66385

Slide-A-Lyzer Dialysis Cassette Equipment

0.1-0.5 ml Equipment

$ 88

3 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description 66380 Slide-A-Lyzer Dialysis Cassette

Product # Description 66003 Slide-A-Lyzer Dialysis Cassette

U.S. Pkg. Dimension Worth 10/pkg. $ 70

12 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description 66012 Slide-A-Lyzer Dialysis Cassette

U.S. Pkg. Dimension Worth 8/pkg. $ 90

30 ml Capability Slide-A-Lyzer Dialysis Cassettes

0.5 ml Capability Slide-A-Lyzer Dialysis Cassettes Product # Description 66383 Slide-A-Lyzer Dialysis Cassette

3 ml Capability Slide-A-Lyzer Dialysis Cassettes

U.S. Capability Pkg. Dimension Worth 0.5-3 ml 10/pkg. $ 73

66381

Slide-A-Lyzer Dialysis Cassette

0.5-3 ml

5×10/pkg. $340

66382

Slide-A-Lyzer Dialysis Cassette Equipment

0.5-3 ml

Equipment

$102

Product # Description 66030 Slide-A-Lyzer Dialysis Cassette

U.S. Pkg. Dimension Worth 6/pkg. $ 72

Pierce Protein Analysis Merchandise Tel: 815-968-0747 or 800-874-3723 • Fax: 815-968-7316 • Buyer Help E-mail: [email protected] © 2007 Thermo Fisher Scientific Inc. All rights reserved. These merchandise are provided for laboratory or manufacturing functions solely. All logos are property of Thermo Fisher Scientific Inc. and its subsidiaries. U.S. patent pending on Zeba Micro Column Know-how. † B-PER Know-how is protected by U.S. patent # 6,174,704. U.S. patent pending on Mitochondria Isolation Equipment and P-PER Reagent Know-how. Slide-A-Lyzer MINI Dialysis Unit know-how is protected by U.S. patent # 6,039,871. Slide-A-Lyzer Dialysis Cassette Know-how is protected by U.S. patent # 5,503,741. Delrin® is a registered trademark of E.I. du Pont de Nemours & Co., Inc. † The RED Gadget is manufactured by Linden Bioscience. Patent pending on RED Gadget by Linden Bioscience. Teflon® is a registered trademark of DuPont.

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