double staining

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From:Cynthia Favara <> (by way of histonet)
To:histonet <>
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	I have been asked to do double labelling. I have worked out
protocols for both procedures using a HRP conjugated secondary and DAB or
AEC. My thinking is I need to switch one procedure to an AP conjugated
system. Since I am just in the thinking stage I have some initial simple
questions and hopes that one with more experience will be albe to point out
potential pitfalls.

	1. What AP substrate works well with DAB? We are hoping for
	2. What is used to quench endogenous alk phos?
	3. Using a dilution grid can I expect to get the same sensitivity
as with the HRP system?
	4.One procedure requires antigen retrieval with pH6.0 citrate. Has
anyone done IHC using DAB then done antigen retrieval?

	And this is just the beginning I look forward to replies meanwhile
I will be hitting the books.

 Thanks in advance,

Cynthia Favara
Rocky Mountain Laboratories
903 S 4th Street
Hamilton, MT 59840
ph: 406-363-9317
FAX: 406-363-9286

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