Silver staining problem
hamburg at sage.cc.purdue.edu
Sat Feb 10 18:18:20 EST 1996
I am analyzing patterns of cell wall proteins by two dimensional
electrophoresis. Unfortunately, the most prominent protein of 23 kDa
(that I am extremely interested in) is not silver stained at all (using
the acidic method).
As I know this protein to be glycosylated, I used the combined Alcian
blue/silver staining method as described by Jay et al. 1990 (Anal.
Biochem. 185: 324-330 - I did not use the PAS-method as I want to stain
both glycosylated and not glycosylated proteins). However, this did not
alter the (one dimensional) pattern of detectable cell wall proteins,
the 23 kDa protein still not being stained.
This protein also shows a strange behavior when I use Coomassie-
staining, as (only) sometimes it does not bind the dye very efficient-
ly. I have not yet figured out the reason for that.
Has anybody found the solution to a similar problem???
Any help/suggestions are greatly appreciated.
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