Production of IgM monomers
mrc7 at cam.ac.uk
Fri May 31 07:32:19 EST 2002
In article <0DE50F4E1F75D411803C00508BE70B2D57BE07 at KSB-GUILDFORD>,
Richard Truran <URL:mailto:RichardTruran at KSBiomedix.com> wrote:
> I am researching the activity of a mouse IgM monoclonal and a mouse IgG,
> they both recognise the same antigen, I want to investigate the effect of
> valency, and so want to be able to break the IgM molecule into the
> monomeric form. Is this possible and if so does anyone know how.
> Richard Truran
You need to look at the papers of Arnold Fienstein from the 70s (He did a
lot of the pioneering work on the structure of IgM). I used a method of his
for my PhD work in 78-81.
Buffer 0.3M NaCl 0.2M TrisHCl pH 8.0 10mM EDTA
(pH8.0 pKa of cysteine) Reduce with 10mM DTT (equiv of 20mM thiol) for 1
hour at room temeperature.
Alkylate with iodoacetamide at a 50% M excess over thiol for 1 hour at
room temperature in the dark.
Dialyse against PBS.
M.R. Clark, PhD. Division of Immunology
Cambridge University, Dept. Pathology
Tennis Court Rd., Cambridge CB2 1QP
Tel.+44 1223 333705 Fax.+44 1223 333875
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