Re: 1st dimension gels

Suming Hua (shua@umaryland.edu)
Wed, 20 May 1998 10:46:58 -0400 (EDT)

Thioglycolate (not thioglycholate) should be as good as mercaptoacetate.
Usually, the gel contains some peroxide , such as APS, which can oxidize
the free amino group to block it. Prerun gel will get rid of all the
oxidizing reagents to protect proteins from becoming blocked.

On Tue, 19 May 1998, Helene Hill wrote:

>
> Would thioglycholate be just as good as mercaptoacetic? I'm new at this:
> what do you mean that the protein is blocked in the 1st dimension gel?
>
> On Tue, 19 May 1998, Suming Hua wrote:
>
> > Prerun gel at 90 volts in your buffer, the uper buffer should contain
> > mercaptoacetic acid, sodium salt (0.2g in 300 ml buffer) or other
> > reducing reagent, for 30 min, then run your sample. The protein should be
> > protected from becoming blocked.
> >
> > On Mon, 18 May 1998, Sandra Smith wrote:
> >
> > > Dear Members,
> > >
> > > We have a user who is wondering what can or should be done to protect
> > > proteins from becoming blocked in the 1st dimension gel. We prerun our 2nd
> > > dimension gels and age them to insure complete acylamide polymerization,
> > > but we are unfamiliar with precautions for the 1st dimension. Any advice
> > > would be greatly appreciated.
> > >
> > > Michelle Gadush and
> > >
> > > Sandie Smith
> > > Protein Microanalysis Facility,
> > > Institute for Cellular and Molecular Biology
> > > University of Texas, Austin, Texas 78712
> > > Email: sandie@mail.utexas.edu
> > > Phone: 512 471-3741, Fax: 512 471-2149
> > >
> > >
> > >
> >
> >
>
> Helene Z Hill, Ph.D. Internet: hill@umdnj.edu
> Head, Section of Cancer Biology Voice/fax: (973)972-3421
> Mail: MSB-E586
> NJ Medical School
> 185 South Orange Ave
> Newark, NJ 07103-2714
>
>