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Doubling labelling with IF with primaries from same host species- how?

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nkm

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HI,
I am trying to do a colocalization with paraffin embedded mouse lung tissue. However, both my primaries are rabbit anti-mouse and i can't find good primaries that arent rabbit.
However, I am worried if both my secondaries are say donkey anti rabbit there may be cross reactivity... is there a way around this without finding different primaries?
thanks in advance

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Posted Jul 05, 2007, 15:30 PM
COHscientist

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I guess you can try and get one of your primary antibodies conjugated with fluorecense instead of using secondary antibody. Otherwise, you will get cross reaction for sure.

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Posted Jul 05, 2007, 16:34 PM
mary-annRSA

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Ever tried chicken antibodies as primary or a secondary? Their easy to induce and isolate providing you have the ethical agreements in place. Speak to the authors of this paper and see whether you would be able to buy some chicken anti-mousse abs from them.

Goldring, J.P.D., and Coetzer, T.H.T. (2003) Isolation of chicken immunoglobulins (IgY) from egg yolk. Biochem Mol Biol Edu 31: 185187.

Best of luck!

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Mary-Ann Chetty
maryannchetty@gmail.com

Posted Oct 09, 2007, 11:43 AM
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