[Histonet] Reply double immunostaining on same species, mouse

[Gayle Callis]

We do double and triple immunfluorescence staining on mouse tissues with all antibodies raised in same host species only on mouse tissues,  rat antiMouse.   It takes some careful blocking and organization, but it works beautifully.  Please contact me and I will discuss the details on how to set up the staining, but supply some details on what you want to see as green or red fluorescence.  This is only done on frozen sections and NOT on formalin fixed paraffin embedded tissue to eliminate autofluorescence. There are several ways to set this up, and we do use some biotinylated primary antibodies along with Molecular Probes Streptavidin Alexa fluor dyes.  Gayle M. CallisHTL/HT/MT(ASCP)Bozeman MT You can still have issues with the second red detection trying to stick to the first AB unless you do some very aggressive blocking.
Patsy

-----Original Message-----
Sent: Saturday, July 12, 2008 9:58 AM
To: histonet
Subject: [Histonet] Double immunostaining with two same source 1st Antibody

Hi all

Just wonder any one of you tried to perform IHC using two primary antibodies from same source, mice, for example?

The process is that 
Mice anti-A (1st IHC)->secondary antibody labeled with GREEN
fluroscence->wash
-> Mice anti-B (2nd IHC)-> Red fluroscence

then there's no cross reaction.

thx.

2008-07-12 



tf 
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