I am trying to help somebody out and would appreciate your input.
I would like to find a membrane that DOES NOT bind protein. The purpose
would be to protect a nitrocellulose membrane during electroblotting. The
problem is that sometimes the gel will stick a little bit to the membrane
after transfer and this results in a lot of background artifacts after
staining. The problem becomes worse as the gel ages. I have been advised
that cooling of the gel during transfer might help but I am told that this
would not be easy to implement.
I suggested that a "spacer" membrane may help. The ideal "spacer" would have
large pores and be very low protein binding. This would allow the protein to
pass through to the nitrocellulose and also stick to the gel instead of the
nitrocellulose. My only problem is that I don't know of anyone who ever
(intentionally) created a membrane in sheet form that does not bind protein
and could be used in a transfer apparatus. Even if we find one, there may
still be problems with band diffusion but it would still be worth a shot.
Any comments/experiences would be greatly appreciated.
Thanks,
Chuck Felice
Purification Technology and Validation
Centocor, Inc.
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This archive was generated by hypermail 2b29 : Wed Nov 21 2001 - 14:17:19 EST