correction for #2--digest in pepsin (0.4% in 0.1N HCl) for 5 min., not 10
min.

> ----------
> From: 	Connolly, Brett
> Sent: 	Wednesday, April 7, 1999 12:07 PM
> To: 	'histonet'
> Subject: 	F4/80 IHC responses
> 
> 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
> 
> 
> Brett M. Connolly, Ph.D.
> Merck Research Laboratories
> Dept. of Human Genetics
> WP26A-3000
> P.O. Box 4
> Sumneytown Pike & Broad St.
> West Point, PA 19486
> ph.215-652-2501
> FAX. 215-652-2075
> email:brett_connolly@merck.com
> 
>