I have been doing B-gal staining on frozen sections without success. I use the SIGMA kit for senescent cells detection, which should be pretty straight forward, but I get no staining on my positive controls.
The protocol I follow is this one:
-Incubate x-gal sol at 37*C for one hour
-Defrost sections for 3 min
-Fix with 1X fixative sol (from kit) for 7 min
-Wash 3 times with 1x pBS
-Add staining mix and incubate overnight, at 37*C
-Countersatin with nuclear red
I don't know if a might be missing something or if my B-gal activity is being affected by a step in the protocol.