Heavy/light chains on Coomassie gel???
mrc7 at cam.ac.uk
Tue Oct 24 07:58:14 EST 2000
In article <Pine.A188.8.131.5210240200050.86132-100000 at acs4.acs.ucalgary.ca>,
Neal Robert Melvin <URL:mailto:nrmelvin at ucalgary.ca> wrote:
> 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??
There are two light chains and two heavy chains in each IgG molecule but
the heavy chains are approximately twice the Mr thus you would expect them
to bind twice as much coomassie blue for the same number of molecules.
o/ \\ // || ,_ o M.R. Clark, PhD. Division of Immunology
<\__,\\ // __o || / /\, Cambridge University, Dept. Pathology
"> || _`\<,_ // \\ \> | Tennis Court Rd., Cambridge CB2 1QP
` || (_)/ (_) // \\ \_ Tel.+44 1223 333705 Fax.+44 1223 333875
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