F4/80 IHC responses

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From:"Connolly, Brett" <brett_connolly@merck.com>
To:"'histonet'" <histonet@Pathology.swmed.edu>
Date:Wed, 07 Apr 1999 13:07:25 -0400

For any interested parties out there I have received the following answers
in response to my query regarding successful clone F4/80 IHC staining of
mouse macrophages on FFPE tissues:

1. Use 1:100 dilution of F4/80 from Cedarlane Labs 90 min. at RT. Secondary
is mouse adsorbed biotinylated anti-rat IgG. No info on pretreatment.

2. Use 10ug/ml of F4/80 from Serotec (MCA497) O/N at 4C. Pretreat with 0.4%
pepsin, 10 min. at 37C.

3. Use 1:10 dilution of F4/80 from Cedarlane Labs O/N 4C. Pretreat with
trypsin 20 min. at 37C. Secondary is Vector biotinylated rabbit anti-rat
1:200, 30. min. followed by Vector ABC Elite 30 min. and DAB 3 min.
Specimens were skin, fixed 2 hr.. in NBF.

4. Pretreat with 0.1% protease XIV, 8 min. at 37C. No other info. Stains
resident and circulating macs (monocytes, I assume), but not macs in
inflammatory lesions.

Please continue to send me your advice and I will post a follow. Thanks to
all who have responded thusfar.


Brett M. Connolly, Ph.D.
Merck Research Laboratories
Dept. of Human Genetics
P.O. Box 4
Sumneytown Pike & Broad St.
West Point, PA 19486
FAX. 215-652-2075

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