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
AP
