Western Blotting of Membrane Proteins in the presence of SDS
Shaun D. Black
SHAUN%JASON.DECNET at relay.the.net
Thu Apr 22 23:07:14 EST 1993
I'd appreciate your experience as to electroblotting of membrane
proteins on PVDF (polyvinyldenedifluoride) membranes. We've tried a number
of conditions but would be curious as to what others have found optimal for
membrane proteins from SDS-PAGE -> PVDF.
1. Buffers: [50mM Tris / 380mM Glycine] vs [25mM Tris / 190mM Gly]
2. SDS : 0% vs 0.1% (or others?)
3. Methanol: 20% required?
4. Voltage / Time: Is there an optimum or do membrane proteins simply
require more time?
5. Is it important to blot right after SDS-PAGE or can gels be soaked
in blotting buffer overnight before electrotransfer?
Thanks in advance. Your thoughts are much appreciated. Cheers. -Shaun
= Shaun D. Black, PhD | Internet: shaun%jason.decnet at relay.the.net =
= Dept. of Biochemistry | Bitnet: shaun%jason.decnet at thenic.bitnet =
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