Dr Engelbert Buxbaum
engelbert_buxbaum at hotmail.com
Thu Feb 16 08:21:55 EST 2006
vtarun.selvan at gmail.com wrote:
> Hi all
> when I stained my leaf extract which contains glycoproteins using PAGE.
> I m not getting any bands.IS there any specific staining solution for
> GLYCOPROTEINS. Usually we use Nacl for proteins staining, but actually
> my extract contains glycoproteins.Suggest me some staining solution for
> the PAGE product.
Presumably you were using KCl, not NaCl for staining, potassium
dodecylsulfate is poorly water soluble, precipitates and makes the gel
turbid. Only in those places where the SDS is bound to proteins does the
gel stay clear. You can use Cu or Zn/Imidazole for a more sensitive
Specific for glycoproteins is the periodic acid/Schiff-reaction (PAS),
but if you do that in the gel the procedure is cumbersome. Better to do
a western blot first, the proteins are then on the surface of the
membrane and the reagents don't have to penetrate the gel. Instead of
Schiffs reagent you can use certain fluorescent dyes like Lucifer Yellow
for even more sensitivity.
If you do electrophoresis of glycoproteins more often, consider doing
eastern blots instead (electrophoresis with CTAB followed by blotting,
Anal. Biochem. 314 (2003) 70-6). The positively charged CTAB results in
much crisper bands of glycoproteins than the negatively charged SDS.
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