[Histonet] F4/80 Ab clone CI:A3-1 not working for foam cells - why?

[Patsy Ruegg]

F4/80 may not pick up as many of the macrophage like cells as some of the
CD68's.  I really like KPMI CD68.
Patsy 


Patsy Ruegg, HT(ASCP)QIHC
IHCtech, LLC
Fitzsimmons BioScience Park
12635 Montview Blvd. Suite 216
Aurora, CO 80010
P-720-859-4060
F-720-859-4110
wk email pruegg <@t> ihctech.net
web site www.ihctech.net
 

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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of GT Hebert
Sent: Friday, August 05, 2005 9:40 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] F4/80 Ab clone CI:A3-1 not working for foam cells - why?

Hello again,
 
I just worked out my F4/80 antibody on mouse tissue.  The mouse tissue,
aortic arch with athrosclerotic lesions came up negative though.  I would
think that the foam cells (macrophages??) would light up as the do when I
stain with CD68 ... but only 1 or 2 cells outside the lesion lit up.  The
Kupffer cells in the liver and the red pulp in the spleen (my + controls)
came out great and the negative isotype controls on all samples are clear.
Any idea why this may be happening?  I have seen this antibody used on a
number of literature papers staining foam cells in AT lesions (same clone -
just different dilutions (1:25, 1:200m etc -- all paraffin) -- and they
showed staining of foam cells.  
 
My tissues are all treated the same: 24 hour fixation in 4%PF, routine
processed and embedded, 4 micron sections, deparaffinized ... and then IHC
(optimal conditions).
 
I'm sort of lost with this one ... I know the antibody works.  Just the
results are not what I expected.  Any information would be greatly
appreciated.
 
Gustave Hebert
Wyeth Cambridge



		
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