Heavy/light chains on Coomassie gel???

Warren Gallin wgallin at gpu.srv.ualberta.ca
Tue Oct 24 10:21:34 EST 2000


Neal,
	The heavy and light chains are there in equimolar amounts, but the
heavy chain is approximately twice as large as the light chain, so there
is twice the mass of heavy chain protein in an equimolar mixture. 
Coomassie staining is roughly proportional to mass, not molar quantity,
hence the heavy chain band stains approximately twice as dark.

Warren Gallin

Neal Robert Melvin wrote:
> 
> Hello,
> 
> I've been using Coomassie Blue staining to get a rough idea of the
> concentration of normal rabbit serum. When I do this, I notice that the
> bands corresponding to the heavy chains stain significantly more intensely
> than the light chain bands... should these be the rough the same intesity
> since one IgG contains 2 heavy chains and two light chains??
> 
> I'm trying to match the concentration of this normal rabbit serum to a
> known amount of immune rabbit serum so that I can perform an
> immunoprecipitation with equivalent amounts of IgG's... which bands should
> I try to match in intensity (between my immune serum and my non-immune
> serum), the heavy chains of each, or the light chains??
> 
> Neal






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