Purified IgG on SDS-PAGE

Student mucineer at iname.com
Mon Oct 9 07:06:33 EST 2000


Dima,

How different can the sizes between the different forms be? (for non-reduced
and reduced)

Thanks.


Dima Klenchin <klenchin at REMOVE_TO_REPLY.facstaff.wisc.edu> wrote in message
news:8rm20f$3qg_006 at doit.wisc.edu...
> "Student" <mucineer at iname.com> wrote:
> :I 've recently run some antibodies (monoclonal) purified from tissue
culture
> :supernatant on a reducing SDS-PAGE gel (Novex) and stained by colloidal
> :blue. On the heavy chain band, I notice that instead of one band, I have
two
> :very close bands, with the larger one fainter than the smaller one. The
> :light chain, on the other hand, is one fine band.
> :
> :Has anyone come across this? The source of my monoclonal was purified by
> :protein A chromatography. Does the concentration I load onto the gels
give
> :this effect? What other possibilities are there?
>
> Glycosylation. Happens all the time. Rabbit polyclonals routinely
> have quite a collection of bands in light chain range; I've seen
> monoclonals giving two distinct light bands, for all I know heavy
> chains are also glycosilated so heterogenity is certainly possible.
>
>         - Dima
>







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