IgM

[Shaun D. Black]

fnunes at pop.nih.gov (Fabio Nunes) wrote on 30 Sep 1993:
> 
> I'm using a monoclonal IgM produced by a company. This antibody labels
> filaments in CV1 and other epithelial cells.  I'm trying to isolate what
> seems to be a new protein. When I run Western blot I obtain 3 bands with
> this antibody. I'm wondering if that result could be a cross reaction. Does
> anyone have experience in isolate proteins using IgM monoclonal antibodies.
> Any help would be greatly appreciated.
> Thanks in advance
> 
> Fabio Nunes
> NIDCD-NIH
> fnunes at pop.nih.gov
> 

In my experience IgM's have always been "stickier" than IgG's.  That is,
IgM has always yielded much more nonspecific binding.  However, that
doesn't mean that IgM's are useless, just that you need to wash blots quite
stringently.  We have used the procedure of Sheng and Schuster; you might
want to consult: BioTechniques 13: 704-708 (1992) and Biochemistry 33: 6945-
6951 (1994).  Hope this helps!  -Shaun
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  = Shaun D. Black, PhD   | Internet address:     shaun at jason.uthct.edu = 
  = Dept. of Biochemistry | University of Texas Health Center, at Tyler = 
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