RE: [Histonet] F4/80 antibody

From:"Favara, Cynthia (NIH/NIAID)"

I have used this antibody successfully in FFPE mouse tissue using
pretreatment with  citrate buffer pH 6.0  120c for 20 minutes. Can't
remember the dil I used I suggest a dilution curve, you can use liver as
F4/80 will stain Kupher cells nicely. I use vector  biotinylated anti-rat,
mouse absorbed, any reliable supplier should work well followed by
streptavidin and AEC as a chromogen.

Relatively pain free!

Cynthia Favara
903 South 4th Street
Hamilton, MT 59840

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-----Original Message-----
From: Angelicola,Lisa [] 
Sent: Monday, August 09, 2004 12:28 PM
Subject: [Histonet] F4/80 antibody

I'm working on a research project that involves demonstrating macrophages in
formalin fixed paraffin embedded mouse skin wounds. The researcher has
purchased the F4/80 rat-anti-mouse antibody from abcam.
Has anyone out there in histoland had any experience working on fixed,
processed tissue with this antibody? The spec sheet states that it will work
with pretreatment of the tissue (protein digestion, which I would like to
avoid if possible since the tissue is susceptible to falling off). Also
where do you purchase your anti-rat secondary from? I would appreciate any
advice anyone might have as far as pretreatment, dilutions, ect.
Thanks in advance,
Lisa Angelicola
Immunohistochemistry Lab
UCONN Health Center
Farmington, Ct.







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