A 2D electrophoresis problem during IEF

[Jean-Etienne Poirrier]

Hello,

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.

Jean-Etienne Poirrier

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
> RJ
> 
>