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Pierce SDS-PAGE Sample Prep Kit


Eliminates band distortions caused by contaminants.

Pierce SDS-PAGE Sample Prep Kit

The Thermo Scientific Pierce SDS-PAGE Sample Prep Kit quickly and easily removes high levels of salts, denaturants, detergents and other buffer components that interfere with polyacrylamide gel electrophoresis of proteins.

The SDS-PAGE Sample Prep Kit provides a simple and effective method for concentrating samples while removing chemicals that interfere with SDS-PAGE analysis, such as acids and bases, detergents, guanidine and ammonium sulfate. The proprietary PAGE-prep Resin binds protein in the presence of dimethylsulfoxide (DMSO). Interfering contaminants are then removed by washing the protein-bound resin in an easy-to-use spin cup format. Proteins are eluted in a buffer compatible with the Pierce BCA Protein Assay, allowing quantitation of a portion of the processed sample. The eluted proteins can then be combined with sample buffer and analyzed on the SDS-PAGE system of choice.

Highlights:

  • Eliminates gel artifacts caused by incompatible contaminants – removes dyes, reducing agents, detergents, sugars, glycerol, guanidine, urea and ammonium sulfate to provide reproducible results by SDS-PAGE analysis
  • Compatible with the BCA protein assay – allows protein quantitation of the processed sample before gel-loading
  • Enriches dilute protein solutions – concentrates protein samples for SDS-PAGE analysis by six-fold in less than 20 minutes
  • Powerful and flexible – process 2 to 350µL of sample containing up to 70µg of protein; recover 75 to 85% of the total protein in 50µL of elution buffer
  • Convenient, complete kit – includes PAGEprep Resin, DMSO, spin cups and sample loading buffer

Includes:

  • PAGEprep resin slurry, DMSO solution, elution buffer, sample buffer, spin cups and collection tubes

Applications:

  • Inclusion bodies solubilized in guanidine•HCl
  • Samples containing low-pH buffers, thiocyanates or urea
  • Proteins precipitated in ammonium sulfate
  • Concentration of dilute protein solutions

Product Details:

The SDS-PAGE Sample Prep Kit uses a unique resin composed of modified diatomaceous earth that binds protein in the presence of DMSO. The simple protocol involves combining 2 to 350µL of sample, containing up to 70µg of protein, with 20µL of the PAGEprep Resin in DMSO (supplied). Proteins in the sample bind to the resin in 3 to 5 minutes, after which the nonbound contaminating chemicals are washed away. Finally, the purified proteins are recovered from the resin in 50µL of Elution Buffer. The recovered protein sample is then ready to mix with the supplied 5X Sample Loading Buffer for gel loading (or another sample loading buffer). The procedure also allows for dilute protein samples to be concentrated up to six-fold, enabling more protein to be loaded per well. In addition, the Thermo Scientific Pierce BCA Protein Assay (Part No. 23225) is compatible with the Elution Buffer and may be used to determine final protein concentration before gel loading.

For reliable SDS-PAGE analysis, protein samples must be prepared in a buffer free of interfering substances and at protein concentrations adequate for analysis. Many interfering substances, which interfere with typical sample buffers for polyacrylamide gel electrophoresis (SDS-PAGE), are difficult to remove by traditional sample preparation methods. For example, protein samples containing 6M guanidine-HCl will precipitate when mixed with Laemmli buffer for SDS-PAGE, causing the sample to run poorly on a gel. Fortunately, samples containing a wide range of interfering chemicals, such as chaotropic agents, detergents, lipids, pH extremes and salts, can be cleaned up within minutes using the SDS-PAGE Sample Prep Kit.

Eliminate band distortion caused by detergent in electrophoresis.
Eliminate band distortion caused by detergents. Detergent-containing membrane (m) and soluble (s) protein fractions resulting from extraction with the Thermo Scientific Mem-PER Kit (Part No. 89826) were separated by SDS-PAGE with and without prior treatment with the SDS-PAGE Sample Prep Kit. Image is a chemiluminescent Western blot for cytochrome oxidase subunit 4 (COX 4). Kit-treated samples remove the detergent that interferes with electrophoresis while retaining relative protein levels of the original samples.

 

Achieve consistent protein recovery in SDS-PAGE sample clean up.
Achieve consistent protein recovery using the Thermo Scientific SDS-PAGE Sample Prep Kit. Pure proteins (60µg) of assorted molecular weights: 30K, 44K, 80K, 86K and 12K and bacterial lysate were processed using the kit. Protein concentrations were determined with the Pierce BCA Protein Assay (Part No. 23225) and reported as percent protein recovered.

 

Effective protein concentration and recovery for SDS-PAGE samples.

Effectively recover and concentrate protein samples for SDS-PAGE. Purified bovine serum albumin (BSA) samples at 0.1mg/mL were electrophoresed and detected with coomassie stain (Part No. 24590).

Lane 1: MW Marker (Part No. 26691).
Lane 2: 10µL (1µg) of BSA sample.
Lane 3: 10µL of BSA sample after treatment with SDS-PAGE Sample Prep Kit.
Lane 4: 200µL of BSA sample after treatment with SDS-PAGE Sample Prep Kit and recovery in 20µL loading buffer. Intensity of Lane 3 relative to Lane 2 indicates good recovery. Intensity of Lane 4 relative to Lane 2 indicates 10-fold concentration of original sample.

 

Effective removal of SDS-PAGE-interfering reagents. Components that were effectively removed from 20µg BSA samples and the associated protein recovery of BSA.
Removed Substance BSA Recovery
Water (Control) 75%
0.5M Sodium Chloride 80%
2M Ammonium Sulfate 76%
20% SDS 75%
10% Triton™ X-100 75%
6M Urea: DMSO (1:3 ratio) 75%
1M Sodium Chloride 75%
6M Urea 74%
10% CHAPS 80%
25% Glycerol 71%
10% OTG 71%
2M Guanidinium•HCl 70%
40% Sucrose 70%

Related Products:

Precast Gels for SDS-PAGE


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
89888 Pierce SDS-PAGE Sample Prep Kit
Formulation: Multi-component kit
Sufficient For: Processing 50 samples (2 to 300μL each) for SDS-PAGE

Kit Contents:
PAGE-prep Resin, 1mL DMSO-slurry
DMSO (dimethylsulfoxide), 27mL
PAGE-prep Elution Buffer, 5mL
Spin Cups (cellulose acetate filter), 50 cups
Collection Tubes, 72 tubes
Lane Marker Non-reducing Sample Buffer (5X), 5mL
50-rxn kit Product Instructions for product #89888 Pierce SDS-PAGE Sample Prep Kit MSDS for product #89888 Pierce SDS-PAGE Sample Prep Kit Certificate of Analysis for product #89888 Pierce SDS-PAGE Sample Prep Kit $164.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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