Polyacrylamide gels shriveling up during transfer - (Mar/04/2008 )
I have been doing western blots and lately my polyacrylamide gels (7.5% Tris-HCL - BIORAD) have been shriveling up during transfer. Subsequently, the proteins have not been effectively transfered. This has not happended before and I have not changed the protocol. I am using the standard tris-glycine transfer buffer (ph 8.3) - can anyone suggest reasons that this may be occuring?
wet or semi-dry transfer?
native or sds page?
if sds, do you have the correct amount of methanol in your transfer buffer? too much could cause shriveling with pore shrinkage (will make it harder to transfer proteins).
if native, did you use methanol? it's not necessary (except to "wet" pvdf membrane prior to transfer).
do you see any transfer at all? did you stain the gel after?
have you checked your power supply? you may be over-heating the transfer.
i have seen gels shrivel up after opening the transfer apparatus after transfer (semi-dry). no effect on transfer.