Urgent question

From:Lynn Gardner <lynn-gardner@uiowa.edu>

Dear Friends,

We are staining for anitbodies in nuede mice. They have been injected with
rat cells and we want to demonstrate the antibody in the rat cells. The
antibody is a rabbit anti-rat antibody. The problem is we get staining in
the tissue but everything is staining. The more worrysome problem is that
the Rabbit IgG negative control is also staining vibrantly positive even
more so than the antibody stained tissue. Does this mean that the rabbit
anti-rat antibody and the Rabbit IgG are cross reacting with the mouse
tissue? or something else?

I block for Hydrogen peroxide, avidin and biotin and use a protein block
that is serum free. I can't figure this one out. Any hints would be most
appreciated!!!

Thanks gang,
Lynn 




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