SDS-PAGE gel sticking - Gel sticking to the membrane (May/23/2005 )
[FONT=Arial]Hi.
I'm trying to do some SDS-PAGE transfers for use in Western Blotting. Every time I remove my transfer cassette, and try to take the membrane out to block overnight, the gel sticks to it really bad. I can't get it off without having to scrape it a little, which I'm pretty sure is not good to do and will smear/ruin my transfer. Any suggestions?
I'm new here, so forgive me if this is a redundant question - I tried to check the back posts first but couldn't find anything useful. Thanks.
I'm trying to do some SDS-PAGE transfers for use in Western Blotting. Every time I remove my transfer cassette, and try to take the membrane out to block overnight, the gel sticks to it really bad. I can't get it off without having to scrape it a little, which I'm pretty sure is not good to do and will smear/ruin my transfer. Any suggestions?
I'm new here, so forgive me if this is a redundant question - I tried to check the back posts first but couldn't find anything useful. Thanks.
Hey,
You cantry one of these (just suggestions)
a) reduce your transfer time,
soak the tansfered blot-stuck gel in transfer blot for sometime after the complete transfer and then try blocking, c) increase the methanol% in your transfer buffer.If you try and something works let me know.
all the best
Amsies,
The only time this happened to me the gel was heatin gup too much during transfer (wet). I discovered the problem (DOW!) was me stupidly grabbing Tris-HCL and NOT Tris-base. As soon as I went back to Tris-base it no longer stuck 
Good luck.
i've noticed that if the upper sheet of paper is not properly hydrated, gels stucks more. alternative to scrapping : incubate your membrane in TBS or PBS (solution of blocking whithout milk.