help:purification of IgM

Dr. Mike Clark mrc7 at cam.ac.uk
Fri Sep 8 11:55:52 EST 1995


In article <42pinu$3q at arcturus.ciril.fr> , jean-francois stoltz
<mailto:gouin at hemato.u-nancy.fr> wrote:
> 
> I need help for two protocols,
> first do anyone known a technique for the purification of human 
> IgM (normal plasma)?
> second, a protocol for the immunochemical quantitation of purified IgM
> by single radial immunodiffusion?
> Thanks
>  
> frederique Gouin, PhD
> Laboratoire d'Hematologie-Hemorheologie, 
> Universite NANCY 1, France
> E-Mail: gouin at hemato.u-nancy.fr

You don't mention how pure you need the Ig.
Two methods I used some 15 years ago when I was working on
rat and mouse IgM monoclonals were
a) a very quick method is to dialyse against low salt buffer
eg water or perhaps 2mM phosphate (IgMs are euglobulins and
precipitate out of solution at low ionic strength)
b) use gel filtration with a very high exclusion Mr. IgMs have
a Mr of just under 1,000,000 and so can be purified in the excluded
volume whilst other serum proteins are included.

I was given both of these methods by Arnold Fienstein who was responsible
for a lot of the pioneering structural work on IgM.


Mike Clark, mrc7 at cam.ac.uk          http://www.path.cam.ac.uk/~mrc7/
--
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