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
exctracts of 10,000,000 cell on SDS PAGE- still nothing. I strongly
in the expertiseof of somebody of you out there, dear colleagues.
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...
More information about the Proteins