help about GST-fusion protein SDS-PAGE

Emir Khatipov khatipovNO at NOuchicago.edu
Thu Aug 29 21:15:10 EST 2002


A colleague of mine was testing his protein of interest for glycosylation by
treating it with glycosidase and running both treated and untreated proteins
on the SDS-PAGE. From what I saw on his gels, the difference in MW between
those two was quite remarkable, about several KDa.
-Emir

"Jen" <jrobertson1 at sympatico.ca> wrote in message
news:roetmu8nlvpmu5n66hfsu2bof649ue6u68 at 4ax.com...
> are you saying that you cleaved your target protein from the GST and
> this target protein runs higher than native?
>
> there is always case where the recombinant protein has different
> glycosylation and phosphorylation levels than in vivo - and these two
> would both have an effect on mobility in a gell - but i don't know
> about a few KDa's worth.....
>
>
>
> On 29 Aug 2002 11:34:27 -0000, fly1234 at yeah.net wrote:
>
> >I expressed a GST-fusion protein in BL21 and purifid it with glutathione
beads. After SDS-PAGE, I was surprised to find that the apparent Molecular
weight of the purified product (in 20mm glutatione buffer) was several KD
larger than that of pre-purified target protein (showed as a bold band) in
cell lysate. Does anybody have the same experience? Could you help me?
> >
> >
> >
> >
>
>http://biowww.net/mynews/tree.php?group_name=bionet_molbio_methds-reagnts&b
egin=0
>





More information about the Methods mailing list