Resolving LMW proteins

Bernard Heymann bheymann at bragg.bio.purdue.edu
Tue Mar 29 17:40:51 EST 1994


In article <1994Mar28.180207.1 at pearl.tufts.edu>, ajayaram at pearl.tufts.edu
wrote:

> 
> Hi there,
> 
> I'm trying to run a 8 kd protein on a SDS get but I'm having some problems
> getting it to show up due to its small size.  I've tried 10, 12 and 15
> percent gels but the but the protein doesn't seem to be there ( the problem is
> obscured by the fact this protein is being made after an induction and the coli 
> proteins interfere with seeing the bottom of the lane where the 8 kd protein
> would presumably run.)  I've tried running the gels without loading dye in the
> sample buffer thinking that the dye front would obscure the 8 kD band.  This
> didn't help.  If anyone has any suggestions about resolving low molecular
> weight proteins please respond by posting in this group or emailing me
> directly.  Thanks.
> 
> Arul
> ajayaram at pearl.tufts.edu

I don't want to discourage you, but I have had some bad experiences with a
13 kd very hydrophobic protein that simply stains poorly with both
coomassie and silver. These days I have a good purification protocol, and I
don't even try and see the protein in cell lysate. Of course it takes time
and requires large amounts of cells, even though the protein is
overproduced. I am currently busy tagging the protein to be able to find it
in gels and cells, which seems to be the only way to speed up detection.

-- 
Bernard Heymann
bheymann at bragg.bio.purdue.edu



More information about the Proteins mailing list