On 26 Jan 1944, I posted :
"Can anybody suggest a practical protocol to purify mouse IgM by affinity
chromatography? Also, is there any better, simpler way?
To date, I've got three replies
>>Shuan D. Black <SHAUN%JASON.DECNET at relay.the.net> suggested :
Sigma (and others, I presume) sells an anti-IgM antibody Sepharose
that can be used to purify IgM. I believe they send directions with the
affinity phase. Hope this can help you.
>>Fred Garbrecht <fred at bmt.mcw.edu> wrote :
I'm not aware of any great affinity method for IgM purification.
Pierce Chemical is marketing some kind of IgM binding resin, but
I have not tried it (seemed very expensive when I looked into it).
Some IgMs may bind to ConA or G, so you may want to try a little
pilot experiment to see if your antibody binds. We purify our
IgMs using a combination of precipitation (ammonium sulfate or
dialysis at low ionic strength), gel filtration, anion exchange, and
hydrophobic affinity columns.
>>Greg Denomme <denomme at fhs.csu.McMaster.CA> who has already post his
answer to the net.
I wish to thanks them for their kind and useful suggestions. Currently
we are also using precipitation by ammonium sulfate followed by G-200
column; but I find these steps to be time consuming.
Dept. of Microbiology, Faculty of Sciences