Steven,
Go to the ABRF web site and search on "2d of membrane proteins" you'll
pull up some excellent discussions on handling membrane proteins, esp on
their introduction into the 2nd dimension,
Deb McMillen
Institute of Molecular Biology
University of Oregon
Eugene OR 97403
On Wed, 15 Nov 2000, Steven White wrote:
> 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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