Immunocytochemistry

[TOBY CANNON]

I have an immuno cytochemuisty problem

I wish to stain fixed cells for two different poteins. I have selective
antibodies to each and one which recognises both. Unfortunately, they ae
all aised in abbit. Is there a simple way of labelling the antibodies
directly, so that I could double-stain, or would omitting a secondary
antibody step lose too much amplification.

Alterrnatively is thee a way to blot sequentially with say anti protein 1
then say FITC-labelled anti-rabbit, then anti prrotein 2 then fluorescein
labelled anti abbit to get a double stain

The only other thing I can think of is to modify each antibody before use,
by covalently adding a different immunogenic prosthesis to each and using
secondaries against the prostheses

Any help gratefully received

toby