Components in this kit are prepared with pure chemicals according to the proprietary technology. To prevent protein degradation, a ready-to-use protease inhibitor cocktail is provided. One Kit is consisted of five reagents enough to enrich four compartment proteins from 5g tissues or about 125 million cells. The efficiency of subcellular fractionation has been investigated by SDS-PAGE and immunoblotting of selected marker proteins. Comparing with C6012-25: CNMCS Compartmental Protein Extraction Kit, BioAssay™, C6012: CNM Compartmental Protein Extraction Kit, BioAssay™ has everything except CS buffer for cytoskeleton protein extraction.
Quality Control
One kit of this lot has been tested to go through the complete compartmental protein extraction procedure from rat colon. Four compartmental proteins from the extraction are used for electrophoresis, transferring to PVDF membrane and immunoblotting with mouse anti-GAPDH, anti-histone H1, anti-vimentin, and Na/K+ ATPase as primary antibodies, and HRP-conjugated anti-mouse IgG as secondary antibody. The separation of four compartment proteins is confirmed by the enrichment of GAPDH in the cytoplasmic fraction, Histone H1 in the nuclear fraction, Na/K+ ATPase in the membrane fraction, and the unique localization of Vimentin in the cytoskeleton fraction.
Kit Components
C6012-25A: Buffer C: HEPES, pH 7.9, MgCl2, KCl, EDTA, sucrose, glycerol, sodium orthovanadate, 1x18ml C6012-25B: Buffer CS: Pipes, pH 6.8, MgCl2, sodium chloride, EDTA, sucrose, SDS, sodium orthovanadate, 1x3ml C6012-25C: Buffer M: HEPES, pH 7.9, MgCl2, KCl, EDTA, sucrose, glycerol, sodium deoxycholate, NP-40, sodium orthovanadate, 1x6ml C6012-25D: Buffer N: HEPES, pH 7.9, MgCl2, sodium chloride, EDTA, glycerol, sodium orthovanadate, 1x6ml C6012-25E: Buffer W: HEPES, pH 7.9, MgCl2, KCl, EDTA, sucrose, glycerol, sodium orthovanadate, 1x50ml C6012-25F: Protease Inhibitors (50X): A cocktail of protease inhibitors, 1x660ul
