IgM purification?

Mike Preston preston at warren.med.harvard.edu
Fri Jul 29 16:46:28 EST 1994


In article <30n6fj$m3m at bigfoot.wustl.edu>, gens at biodec (scott gens) wrote:

> I have a 200 uL of a monoclonal mouse IgM antibody in ascites fluid that I 
> would like to purify. This is all I have so I need to get it right on the
> first try. 
> 
> Protein A supposedly binds to IgG and IgM, but most catalogs present 
> tables which state that their Protein A columns do not bind mouse IgM. Is 
> mouse an exception, then? Why the apparent contradiction?
> 
 Thanks in advance
> 
> Scott Gens
> gens at biodec.wustl.edu

Scott, 
gel filtration seems to be the best way to purify IgM.  Try G-200 or S-300
from Pharmacia (Bio-Rad also makes a good gel for this purpose but I can't
remember the number).  Your sample will be diluted about 5 fold after gel
filtration but you are assured virtually 100% recovery. You can concentrate
if needed afterwards but at appx 1 mg/ml of antibody in ascites you prbably
will not need to.  For more details on the methods try your library...
should have some books on monoclonals or immunotechniques.
Good luck

Mike Preston

preston at warren.med.harvard.edu
617-432-0152



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