Re: IHC on Mouse cell cultures

From:Greg Dobbin <dobbin@Upei.CA> (by way of histonet)

If you have a decent amount of the polyclonal antibody you could try
adsorbing it on a mouse cell culture that does NOT have the target
antigen you are staining for. I don't have a specific protocol for you,
but you should be able to find one in most cell culture or virology text
books. Briefly, you incubate the antisera on a washed cell culture at
37 C, rocking the flask periodically to insure maximum adsorption
of non-specific antibodies.
If you only have a small amount of the polyclonal, you may try adding
normal mouse serum to your antibody dilution buffer. Anywhere
from 0.5 to 5% normal mouse serum in the dilution buffer may
"quench" the nonspecific reactants.
Maybe someone else could comment on these suggestions and/or
expand on them. Good luck. Greg

Date sent:      	Thu, 31 Aug 2000 13:31:31 -0500
From:           	Colleen Forster <>
Subject:        	IHC on Mouse cell cultures
Forwarded to:

> Hello Histonetters,
> I am trying to do IHC on mouse CNS cell culture slides. If I
> do monoclonal antibodies the results are great! However, when I use
> polyclonals either goat or rabbit the bkgd is exceptionally high.
> Could some of you please share some suggestions on cleaning up
> this problem??
> As always, Thanks in advance.
> -- Colleen Forster
> U of MN, Dept. of Neurology
> 612-626-2477

Greg Dobbin
Pathology Lab
Atlantic Veterinary College, U.P.E.I.
550 Unviversity Ave.
Charlottetown, P.E.I.
Canada,  C1A 4P3
Phone: (902)566-0744
Fax: (902)566-0851

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