[Histonet] RE: IHC double-labeling -- same secondary?
C.M. van der Loos
c.m.vanderloos <@t> amc.uva.nl
Wed Mar 29 13:29:43 CST 2006
Hi Anna Elisse,
To my opinion there is a strong preference of using second step
reagents of the same host. That makes it very safe to mix them in a
cocktail (time saving!) without any chance of unexpected
cross-reactions. In your case you could mix donkey anti-rat/FLUO +
donkey anti-goat/biotin. Don't forget that you could mix your
primaries in a cocktail as well!
Chris van der Loos, PhD
Dept. of Pathology
Academic Medical Center M2-230
NL-1105 AZ Amsterdam
Date: Wed, 29 Mar 2006 10:52:59 -0500 (EST)
From: "Anna Elisse Beaudin" <aep10 <@t> cornell.edu>
Subject: [Histonet] IHC double-labeling -- same secondary?
To: Histonet <@t> lists.utsouthwestern.edu
I am performing double labeling IHC on mouse brain tissue with two
primaries from different species, one raised in rat, the other in
I was wondering if it was kosher (or even preferred) to use both a
donkey anti-rat secondary and a donkey-anti goat secondary. if so,
I incubate with both together as a cocktail? my donkey anti-rat is
flourophore conjugated, and I was planning to use biotinylated donkey
anti-goat followed with a flourophore-conjugated streptavidin. any
suggestions/comments would be greatly appreciated.
Thanks in advance,
Division of Nutritional Sciences
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