help for IHc stains on whole mount embryos

From:Colleen Forster <cforster@tc.umn.edu>

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
antibody
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
612-626-2477
cforster@tc.umn.edu





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