Hello all -
A collaborator comments on the challenge of membrane proteins on 2D gels:
Right now, I
found the right conditions (detergents and others) for membrane proteins
solubilization, I got really nice IEF gels. The problem is that most of the
proteins (about 90%) do not transfert from the first dimension (the IPG
strip from Biorad) to the second dimension (SDS-PAGE). It seems like their
are interactions (maybe hydrophobic) between the IPG strip matrix and the
membrane proteins. I used TBP as a reducing agent and I am wondering if
this is not the problem. I am right now thrying several controls with
either DTT or TBP as reducing agents. I hope I will solve this problem
soon. This is extremely time consuming. Any idea?
Can anyone comment on the above situation?
Thanks,
Dr. Steven P. White
Oklahoma State University
Department of Biochemistry & Molecular Biology
246 NRC
Stillwater, OK 74078
Phone & Voice Mail (405) 744-6191
Fax (405) 744-7799
e-mail swhite@biochem.okstate.edu
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