Purified IgG on SDS-PAGE

Dima Klenchin klenchin at REMOVE_TO_REPLY.facstaff.wisc.edu
Fri Oct 6 21:32:47 EST 2000

"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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