Could this liver homogenate be used for Western Blot? - (Feb/03/2009 )
Hi, I am totally new in this field. My lab usually uses 0.25M sucrose and 0.1mM EDTA to homogenize fresh livers from rat. I just wonder if this homogenate could be used for Western Blot. If not, how much tissue (weight) will be needed for it? Thanks!
there will be a lot of proteins in a homogenate, your protein of interest needs to be in enough abundance that it can be picked out from the crowd (depends on sensitivity of antibody).
I guess you should homogenize in ordinary lysis buffer with protease and phosphatase inhibitors on ice. It is better to homogenize large piece to be representative as some area may be fibrotic or cirrhotic .