Dear Nora:  Mary had some specific questions on her samples but I can
give you the same reference I gave her.  You need two different
immunhistochemical systems and two different chromogens.  You can do
sequential staining or simultaneous double staining depending on your
choice of systems, abs, etc..  Color contrast is important.  My friend,
Dr. Chris M Van der Loos in Amsterdam has done a lot of work with double
and triple staining.  He has just finished writing up a manual for DAKO,
Immunoenzymatic Double Staining Methods, which I found to be very
helpful for those wanting to double stain things.  I would highly
recommend this as a good starting place.  I assume the manual's free
from your DAKO rep, right Christopher?  The usual disclaimers: I don't
work for DAKO and you don't have to use DAKO products to make it work.
Also double staining isn't necessarily more helpful for any "diagnostic"
uses than staining two separate slides but may help in research work and
looks great!
Regards, Tom Kuwahara

Nora Fox wrote:
>
> I would like to see this too--as may lots of others.
>
> Thank you to all of you who have responded to my many questions.
>
> Nora
>
> >
> >>Good Morning,
> >>        Would someone who has some experience with double labeling[IHC]
> >>please e-mail me directly? I have a few questions from one of my
> >>colleagues...we're interested in one Ab w/DAB and one with alk.phos. Thanks
> >>                              Best Regards,
> >>                              Mary Vaughan HT(ASCP)
> >>                              Roswell Park Cancer Institute
> >>                              Pharmacology + Therapeutics
> >>                              Elm + Carlton Sts.  CDC-121
> >>                              Buffalo, NY 14263
> >
> >
> >
> Nora Fox
> Division of Orthopaedic Surgery
> Stanford University
> Edwards R144, MC 5341
> 300 Pasteur Drive
> Stanford, CA 94305
> phone: (650)725-2746
> fax:  (650)723-6396

--
*******************************
Thomas J. Kuwahara
Senior Immunohistochemist
Advanced Pathology Systems
3801 Sacramento St. suite 621
San Francisco, CA 94118
415 750 6800 x23067 tel
415 750 2332 fax
tom@adpath.com