Cheryl,
I have done eNOS/NOS Type III on rat vessels. The primary was  from
Transduction Labs. I didn't need to do any AR, but it was crucial to make up
the antibodies in PBS/ 1% BSA/with calcium and magnesium. I made my life
easy and "procured" a bottle of premade PBS / Ca & Mg  from the tissue
culture room :) I used primary dilution of 1:500 overnight at 4C, secondary
1:500 for 60 minutes at RT.
Good luck, if you would like a copy of my complete protocol let me know and
I'll forward it on to you.

Mary Georger
Center for Cardiovascular Research
University of Rochester Medical Center
KMRB Room 2-9816
601 Elmwood Avenue.
PO Box 679
Rochester, New York  14642
585-273-1548

> ----------
> From: 	Cheryl Crowder
> Sent: 	Thursday, October 31, 2002 10:26 AM
> To: 	histonet
> Subject: 	eNOS in eyes  and Sihler stain
> 
> Good morning - I saw you question on the Histonet about a stain for nerve
> branching.  I have found the directions and will send them to you by 
> personal e-mail. 
>      Now, could you help me with my problem?  I am starting a research 
> project using eNOS I, II, III.  The primary Abs are from Serotec and 
> their help is minimal.
>     Has anyonedone this type of IHC and what procedure did you use?  The 
> scant info I got did not mention antigen retrival and had a broad area 
> for dilutions.  Any help you can give me would "make my day".  Thank 
> you.  Cheryl
> 
> 
> Cheryl Crowder, BA, HTL(ASCP)
> Chief Technologist
> Department of Pathobiological Sciences
> School of Veterinary Medicine
> Louisiana State University
> Baton Rouge, LA #160#70803
> 
> (225) 578-9784
> FAX #160#(225) 578-9720
> 
> 
>