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[Protein-analysis] Re: on PAS for glycoprotein

jh via proteins%40net.bio.net (by biocjh from gmail.com)
Wed Oct 15 06:34:36 EST 2008


Thanks Dr Engelbert Buxbaum
I want to know how sensitive the PAS method is? If I want a
glycoprotein visulized on blot, how much sample should I load in the
gel?

2008/10/15 Dr Engelbert Buxbaum <engelbert_buxbaum from hotmail.com>:
> Am 04.10.2008, 07:57 Uhr, schrieb jh <biocjh from gmail.com>:
>
>> Dear all,
>>
>> I stained a gel with PAS(periodic acid stain) method for visualizing
>> glycoproteins.
>> However, even protein markers and the reconstructed protein expressed
>> by E. coli could be stained.
>> Whether the PAS method is specific for visualizing glycoproteins?
>> And how to explain the non-glycoprotein can also be stained?
>>
>> I am appreciated of any help of you!
>
> Many marker proteins are glycosylated, this is true for almost all proteins
> that are secreted.
>
> Since you did not say how you did the staining here a reference that I know
> works:
>
> Segrest,J.P. & Jackson,R.L.: Molecular Weight Determination of Glycoproteins
> by Polyacrylamide Gel Electrophoresis in Sodium Dodecyl Sulfate
> Meth. Enzymol. 28 (1972) 54-63
>
> G. Fairbanks, T.L. Steck & D.F.H. Wallach: Electrophoretic analysis of the
> major polypeptides of the human erythrocyte membrane
> Biochemistry 10 (1971) 2606-2617
>
> Btw: It is much more convenient to do the PAS-staining on a western blot
> than in the gel, the incubation times are shorter and less reagent is
> required.
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-- 
Cao Jianhao

Department of Biochemistry
School of Lifesciences
Sun Yat-sen Uniersity
Xingang Road, west
Guangzhou, 510275
CHINA



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