protein extraction - (Dec/21/2007 )
Im looking for a protocol to extract proteins from rat liver tissue. I will then be looking at the level of the nuclear transcription factor ERalpha and ERbeta, (estrogen receptor alpha and beta) by western blotting.
I found a protocol with Pierce online regarding their T-per reagent for total protein extraction but im not sure if this is suitable or should i be using a specific nuclear protein extraction protocol such as the one from Qiagen. Or does anybody have an in-house method?
Im just not sure how to go about choosing a buffer, the salts, protease inhibitors etc, specifically needed for this tissue and the protein im looking at, for making an in-house procedure.
I look at GR, PR and MR and use an AllPrep kit from Qiagen to isolate RNA, DNA and protein from the one tissue source. It works. The standard method with these NHRs is the T-Per reagent from Pierce so it should work with ERs.
Hope this helps,