Re: Double Staining

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From:Mary Vaughan <>
To:"Cindy Chard-Bergstrom" <>
Date:Tue, 27 Apr 1999 13:51:41 -0500
Content-Type:text/plain; charset="us-ascii"

Hi Cindy,
        I've done double staining on paraffin sections. A few points to
        1) You should have the whole method for each antibody worked out
separately before you try to do them in tandem.
        2) If one requires pretreatment and one does not, do that one first,
and then the one that does require a pretreatment second.
        3) I've done both with HRP - I used DAB on the first procedure, and
DAB with Nickel Chloride [looks black instead of brown] for the second. In
the assay that I did, one of the Ab was cytoplasmic, and one was nuclear.
        If you have any specific questions, feel free to e-mail me.
				Best Regards,
				Mary Vaughan HT(ASCP)
				Roswell Park Cancer Institute
				Pharmacology + Therapeutics
				Elm + Carlton Sts.  CDC-121
				Buffalo, NY 14263

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