multimer on denatured SDS-PAGE?

Rick00100 rick00100 at aol.com
Sat May 2 21:55:57 EST 1998


Hi everyone,

I'm a newbie in protein work and Western, and recently I'm trying to raise
antibodies to my protein, which is about 22kd.  I got some polyclonal Ab, and
when I do a Western with CHO cells-transfected protein, I would not see a band
with one of my Ab.  However, after deglycosylating the cell lysates, I can see
a band at around 40kd, and a faint band at 22kd.  I'll try to do the
appropriate peptide competition experiments when I get a hold of some peptides.
 At the mean time, can anybody tell my whether that 40kd band can be a dimer of
my protein?  I thought protein multimers (homo- or hetero-) would dissociate in
a reducing condition (5% b-ME, 5% SDS + boiling 10 minutes).

Thanks for the advice

Ricky, JHMI



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