A 2D electrophoresis problem during IEF
jepoirrier at nospam.student.ulg.ac.be
Fri Mar 8 09:57:33 EST 2002
I use urea, thiourea, AG501-X8, IPG Buffer 3-10, DTT and Chaps. It works
fine. Do you rehydrate your gels correctly ?
Very stupid question but it may happen : how do you see your bands ? Do you
reveal your 1D gels ?
Because I never see bands when 1st dimension is finished. But I sometimes
reveal my 1st dimension gels with my 2nd dimension gel AFTER the transfer
and migration in the 2D gel. But I use a fix solution -> I cannot get any
protein out of fixed gel.
It's arather stupid question, I know.
Hope you'll find some help.
RJ <RJ230 at hotmail.com> wrote in news:3C771F04.37457ED9 at hotmail.com:
> I hope I can get an answer to this question. After the isoelectric
> focusing step is finished successfully, most of my proteins seem to
> precipitate in the IPG strip as I can easily see them as nicely focused
> bands throughout the strip. The problem is to get the proteins move into
> the second dimension gel. Has any one come across such a problem and if
> yes, is there a solution? I should mention that my protein sample
> contains glycoproteins, some of which are highly glycosylated. My
> rehydration buffer system is Urea/Thiourea, SB3-10, CHAPS, Biolyte 3-10.
> Thanks for your help
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