I have used Joanne's idea before, and it has worked quite well. the only
problem I have observed is that the first chromogen sometimes looses it's
"crispiness". So I would suggest that you be careful which enzyme/chromogen
combination you use first. In my experience I have found that peroxidase/DAB
is the least affected while alk-phosp/nbt-bcip is the worse.

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu
[mailto:histonet-bounces@lists.utsouthwestern.edu]On Behalf Of Joanne
Mauger
Sent: Wednesday, September 22, 2004 8:03 AM
To: histonet@lists.utsouthwestern.edu; Inga.Hansson@neuro.uu.se
Subject: Re: [Histonet] double stainings


Dear Inga,
For double staining such markers, you can first stain the antibody that does
not require heat. After the chromogen step, do your heat retrieval, and then
stain with the next primary Ab.
Jo

>>> "Inga Hansson"  09/22/04 07:05AM >>>
Hi everyone!

If I want to do a double staining using one antibody that requires
HIER and another that will stain everything if heated........what do
I do?
Can anyone help me?

Thanks !

Inga
--


Inga Hansson
dept. neuroscience, div. neurobiology
PO Box 587
Biomedical Centre
Husargatan 3
S-751 23
Uppsala
SWEDEN

phone:+46-18-4714384
fax: +46-18-559017

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