Double labelling immunohist. with 2 mouse ab.s

[Dave Peritt]

In article <4utku1$hib at alfa.physiol.su.oz.au> Keith Brain,
keithb at physiol.su.oz.au writes:
>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


NO!!!!!!
everything will bind to everything leading to a big mess...  You need to
directly label with FITC or biotin.  You may also use isotype specific
Abs.  I suggest FITC labeling one and the other biotinylate.  Both simple
procedures.  SA any color you want.....