Purification of IgM & C4 from human serum

Al awdodds at molbiol.ox.ac.uk
Wed Oct 9 14:43:04 EST 1996


In article <325591AA.71CA at mr.net>, Library <library at mr.net> writes:
> Hello-
> I am looking for any ideas on how to extract (not necessarily purify)
> either IgM or C4 (Complement 4) from raw human serum.  We are exploring
> ways of elevating either protein in calibrator products and are looking
> for any helpful suggestions.  Thanks.
> -Janene Lenard
>  Protein Chemist

Do a euglobulin precipitation at pH 7.4 (I vol serum + 4 vol 5mM CaCl2 
at 4 degrees for two hours, spin and remove precipitate.  Then adjust pH to 5.5 
and leave for another 15 min then spin.  The first pecipitate will contain most 
of the IgM (plus C1 and C4bBP) the sercond pecipitate will contain most of the 
C4 (Gigli et al 1977 Biochem J 165, 439)  

Alister 




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