RE: double IHC/in situ #2 (Paul Klosen)

>We are trying to establish IHC/ISH procedure in our lab, and it looks like
>when I do antigen retrieval, the ISH probes tend to bind non-specifically.
>What is the sequence of your work? when do you perform the retrieval step?
>It would be great if you could e-mail your protocol for the pituitary
>Thanks a lot in advance,
>Anna Logvinova, M.D.
>Morphology Core Supervisor
>Buck Institute
>8001 Redwood Blvd
>Novato, CA 94948

I forgot to mention that we tried a pretreatment technique for ISH
published in JHC (don't remember the ref though) which called on a
microwave treatment instead of proteinase K treatment. This technique did
not improe our ISH signals but did clearly cause non-spcific binding of all
of our probes to the tissue, and this whether used on paraffin, PEG or


                         (o -) O
Paul Klosen, PhD
CNRS UMR 7518 Neurobiologie des Fonctions Rythmiques et Saisonnieres
Universite Louis Pasteur
12 rue de l'Universite
F-67000 Strasbourg, FRANCE
tel:    fax:

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