IHC - rinsing after blocking/permeabilizing sample? - (Sep/20/2011 )
I have a question concerning IHC. After I simultaneously block and permeabilize my samples with 1% BSA and 0.3% Trit-X-100 prior to antibody incubation, should I wash the samples several times before primary antibody incubation? I have read somewhere that Trit-X as a detergent can change the final level of antibodies binding to epitopes. Do you guys happen to know what is the best course of action after blocking+permeabilizing? Thanks.
I give them a rinse in PBS with 0.5% tween-20 or blocking solution before adding antibody dilutions.