Formalin Fixed Tissue westerns - (Jun/24/2004 )
Does anyone have a protocol for isolating protein from Formalin Fixed Tissue (not paraffin embedded) for a subsequent western blot. I have been using a few different methods to extract the protein from the fixed tissue resulting in a positive protein assay (coomassie) but when the gels are loaded against control tissue (fresh frozen) no protein runs in the lanes for the fixed while the fresh frozen generates strong bands. Any suggestions?
Formalin fixing and protein analyses donot go hand in hand very well, too many random condensation reactions and cross-linkings are initiated. To achieve better results, formalin fixed tissues must be extracted under nitrogen or argon.