2D PAGE problem

Anton Posch posch at bgfa.ruhr-uni-bochum.de
Tue Jan 13 15:00:14 EST 1998

On Sat, 10 Jan 1998 23:21:38 -0600, p.bayliss at utoronto.ca wrote:

>  When I run the first dimension of a 2D gel on this particular sample,
>something stacks inside the tube (visible by bromophenol blue not getting
>through it).  If left long enough (eg. 4hr) and with less sample, it
>eventually runs out of the bottom of the tube.  It's a nuisance, and may
>be taking some of my protein with it (especially LMW), and it seems to
>temporarily shift the pI gradient (bromophenol blue which DOES get past
>turns yellow, even if it's not near the bottom of the tube).  With silver
>stain, it doesn't stain like normal protein, but stains a very light
>blue.  I haven't stained with coomassie.  In the second dimension, it
>again all runs to more or less the same spot.  I'm thinking it's lipid,
>but I'm not sure.  I've tried acetone and also chloroform extraction with
>minimal success.  I'd like to rid myself of it altogether.
>  Has anyone had similar problems, or have any suggestions?  Anyone know
>of a really good way to solubilize proteins and then get rid of lipids?
>     Thanks in advance for your help!!
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Hi 2D-user,
I´m working very long with 2D technology, but to give you any advice
you have to tell me what sample you work with and how is your sample
prepared for IEF


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