Bio Rad protein assay for liver homogenate - (Jun/27/2011 )
So I'm new to eukaryotes and I need to do a protein assay on homogenized liver tissue. The BioRad assay kit says that the protein needs to be solubilized and that I can't do the assay on a suspension or unfiltered homogenate. Any suggestions on what I'm supposed to do? Thanks!
Basically you need to get the liver into solution - if you have already homogenized the liver, you are half-way there... presumably the homogenate was prepared in a lysis buffer containing some detergents (SDS, triton X-100, tween 20 or 80, NP-40, nonidet p40...) and protease inhibitors. If so, leave the homogenate to sit on ice for 30 min-1 hour and then spin down the un-dissolved stuff at max-speed in a centrifuge (usually done in a small benchtop centrfuge at 13,000-16,000 RCF for 5 min). The supernate is the stuff you want for your assay.