Hi Akhtar, The Bradford assay can only be used if your samples have very low concentration of detergents (here is a table<http://www.sigmaaldrich.com/content/dam/sigma-aldrich/life-science/proteomics-and-protein/migrationpep1/compatibility-chart.Par.0001.Image.-1.-1.1.gif> showing how much detergent can be used). You should use the BCA assay instead: http://www.piercenet.com/browse.cfm?fldID=02020101 (this is the kit that I use) http://www.sigmaaldrich.com/catalog/product/sigma/bca1?lang=en®ion=US
Cheers, Irit On Mar 7, 2013, at 1:40 AM, akhatr hussain wrote: Dear sir/madam I have extracted protein the kidney of mice with RIPA Buffer. Now I want to do the protein concentration estimation of these samples. But my sample is dissolced in RIPA buffer and this does not work accurate with bradford assay as When i dissolved 10 ul RIPA or 5 ul RIPA is bradford solution(from Biorad) is gives colour, though it does not havemy protein protein sample from mice kidney in that. I want to accurately estimate the concentration of the protein in my samples and then to procced with proteomic analysis of these samples for western blot and 2-D gel. May you could kindly guide me what should I do? Thank you best regards, akhtar hussain _______________________________________________ Methods mailing list [email protected]<mailto:[email protected]> http://www.bio.net/biomail/listinfo/methods _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
