protein purification

Thomas Urbig at biokemi.su.se
Tue Jul 22 09:58:54 EST 1997


In article <33D36EB8.68AA at sciborg.uwaterloo.ca>, Wildwind
<swvong at sciborg.uwaterloo.ca> wrote:

> Hello,
> 
> I have extracted protein for 1-D electrophoresis in plant tissue and 
> have had no problems in the past. However, recently (in seed tissue) I 
> have had severe streaking throughout the entire gels (not just in the 
> lanes) following staining, making detection of bands impossible. I asked 
> several co-workers who believe it may be polysaccharides in the sample 
> buffer. Does anyone have any suggestions on how to fix this problem? It 
> would be appreciated.
> 
> Thanks

I would agree on the polysaccaride theory. If you´re working with the
membrane fraction, make a centrifugation an resuspend only the top of the
pellet leaving the whitish lower pellet. Sometimes you have to repeat this
washing step. It helps if you use the second step a small bore tube. If
you are working with the soluble fraction just make a high speed
centrifugation, that should do it.
Hope that helps

Thomas

-- 
Thomas Urbig
Urbig at biokemi.su.se



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