multimer on denatured SDS-PAGE?

rdudley at nrc.uab.edu rdudley at nrc.uab.edu
Sun May 3 02:57:44 EST 1998


In article <1998050302555700.WAA08931 at ladder01.news.aol.com>#1/1,
  rick00100 at aol.com (Rick00100) wrote:
>
> 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

Actually, a number of receptors form noncovalent multimers that don't
dissociate when boiled in BME or DTT and run on SDS-PAGE.  Presents quite a
problem for us.

rich


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