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.
Features of the SDS-PAGE Sample Prep Kit:
• 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 350 µL of sample containing up to 70 µg of protein; recover 75 to 85% of the total protein in 50 µL of elution buffer • Convenient, complete kit—includes PAGEprep Resin, DMSO, spin cups and sample loading buffer
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.
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
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 350 µL of sample, containing up to 70 µg of protein, with 20 µL 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 50 µL 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.