Double labelling immunohist. with 2 mouse ab.s

Keith Brain keithb at
Wed Aug 14 17:45:21 EST 1996

We trying to use immunohistochemistry with two antibodies. 
Unfortunately,  both of these were raised in mice.  Our secondary
antibodies are rabbit anti-mouse.  Could someone point me in the
direction of a suitable protocol for labelling with minimal cross-talk?

We have considered the following protocol;
1.Antibody 1
2. Wash
3. Fluroescein conjugated rabbit anti-mouse (at high concentration)
4. Wash
5. Antibody 2
6. Wash
6. Rhodamine conjugated rabbit anti-mouse

Will this protocol work, is there an easier way, or is it a hopeless
Thanks for you help,
        Keith Brain
        keithb at

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