Fw: help for IHc stains on whole mount embryos

From:Jan Shivers <shive003@maroon.tc.umn.edu>

I would assume that his secondary goat anti-mouse is binding to the mouse
tissue in general (circulating serum immunoglobulins, plasma cells, etc.,
etc.), and not specifically to the mouse IgG primary antibody.  I don't
stain mouse tissue, but I know there are kits out there for use on problems
such as this.   As to the question about whole mount tissue, don't know the
answer to that one.

Jan Shivers
Univ. of Minnesota
College of Veterinary Medicine
Veterinary Diagnostic Laboratory

-----Original Message-----
From: Colleen Forster <cforster@tc.umn.edu>
To: histonet@pathology.swmed.edu <histonet@pathology.swmed.edu>
Date: Thursday, November 30, 2000 4:05 PM
Subject: help for IHc stains on whole mount embryos

>IHC Gurus,
>I am asking for a college for help in staining IHC on whole mount mouse
>embryos. here are the problems:
>The staining appears non specific due to the high background. so is the
>not penetrating the skin or is his block not working?
>He has been using detergents in his buffers and digesting with
>enzymes.The tissues is
>dropped  fixed in methanol/DMSO at RT overnight, rinsed well and placed
>into buffer.
>The block he has been using is 80%fetal bovine serum, the primary is a
>mouse monoclonal.
>The secondary he uses is a goat anti-mouse. The incubation time for the
>primary and secondary
>is 4 C overnight.
>Any suggestions ill be greatly appreciated. Again thanks in advance!!
>-- Colleen Forster
>U of MN, Dept. of Neurology

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