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
