bitter sugar

TandRnawm tandrnawm at aol.com
Sun Sep 18 07:52:01 EST 1994


In article <335eu2$ieq at mserv1.dl.ac.uk>, <SPAGNOL at PLAUTO.CSATA.IT> writes:

I cloned a glycoprotein in a cell line and have antibodies against its
sugar moiety. I do immunofluorescence and the protein is there, then I
perform an immunoblot with the same Ab and nothing stains. I arrived to
load
exctracts of 10,000,000 cell on SDS PAGE- still nothing. I strongly
confide
in the expertiseof  of somebody of you out there, dear colleagues.
Sincerely
Giorgio

Giorgio:
The cloned protein may bear sugars unrelated to those to which the Ab was
developed.  Glycosylation varies by tissue (*within an organism*), culture
conditions, cell line, species, etc. If the protein is mammalian, and
you've expressed it in a non-mammalian system, one would expect the
glycans to be *very* different.  Even if you're using a mammalian
expression system, they may be different.  Of course, the protein may also
be non-glycosylated in your clone...  
Good luck!
Todd Morrill



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