RE: [Histonet] CD68 in FFPE mouse tissue

From:"Elizabeth Chlipala"


We use the same antibodies on mouse aortic root sections and I have
never had a problem with either of them.  We perform the immunostains on
frozen sections.  We follow Gayles protocol for sectioning and fixation,
air dry overnight and then fix in a mixture of acetone/ethanol and then
proced with staining.  The stains always turn out great.  My protocol
might be a bit different I use dako's rabbit anti-rat for my secondary
and then I use their rabbit envision HRP.  I can send you some images if
you want to see how they work for us.  I have not tried the CD68 on
paraffin sections yet but the F4/80 works great on 10% NBF fixed
paraffin sections


Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
P.O. Box 18592
Boulder, Colorado 80308
Office: (303) 735-5001
Fax: (303) 735-3540
Ship to Address:
Premier Laboratory
University of Colorado
MCDB, Room A3B40
Boulder, Colorado 80309

-----Original Message-----
[] On Behalf Of Einar
Sent: Wednesday, June 28, 2006 6:49 AM
Subject: [Histonet] CD68 in FFPE mouse tissue

I am having problems with IHC staining with macrophage-markers in 
formalin-fixed paraffin-embedded (FFPE) mouse arotic root sections.
(Serotec) works well in spleen (after proteinase K digestion) but is 
inconsistent in aortas. I have also tried CD68 clone FA11 (Serotec) with

proteinase K digestion and HIER (citrate, borate, Tris-HCl)  with no 
success. Any tips?
Regards, Einar.
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