Until last year, I had successfully used Transduction's iNOS without any
retrieval with good results.  The lot which I received in 1999 no longer
worked even with various retrieval methods (mouse skin, formalin-fixed less
than 24 hours, paraffin-embedded). I am currently trying a Santa Cruz
antibody and will now definitely give the Upstate Biotech antibody a try
also..thanks Jamie

Mary Ross
Research Associate
Molec Virol, Immuno, Med Genetics
Ohio State University


>Date: 19 Apr 2000 12:24:31 -0500
>From: "Jamie Erickson" <JErickson@genetics.com>
>Subject: Re: iNOS
>
>Kathy,
>            I've used a iNOS antibody (Ab) Rabbit polyclonal (iNOS) Upstate
>biotechnology Cat 06-295 on mouse and Rat tissue on  formalin fixed paraffin
>embedded tissue (FFPE) as I remind all my PI principal investigator I will
>tell you that I only fix for samples for IHC for 24 hour then I process the
>tissue. With some Ab it doesn't matter but this one I've seen problems with
>longer fixed samples. With this Ab I was fortunate to have a good positive
>control Leishmania infected  mouse footpads very positive. So here is what I
>did,  sectioned at 4-5 um air dried overnight placed slides in 60 degree C
>oven for about 20-30 minutes Just to melt the paraffin , and make sure my
>slides are dry) then dewax/ dehydrate to water enzyme digested for 20 minutes
>at 37 w/ trypsin (Zymed labs) then do IHC.
>  Primary Ab (2.5 ug/mL) 1 hour, serum block (5%  goat serum that ever
>secondary is made in), secondary biotinylated Vector labs (anti-Rabbit) . And
>I got great Macrophages infected with parasite staining. I used AEC for
>publication stuff  better contrast but DAB for everything else. There is this
>other chromogen I use NOVARED Vector Labs you can dehydrate to xylene and
>mount w/ permount I use that now...Hope it works for you.



>>> kwalters <kwalters@blue.weeg.uiowa.edu> 04/19 8:33 AM >>>
>Help....I need to do an immunocytochemistry run for inducible nitric oxide
>synthase.  The paper I have uses anti-iNOS (polyclonal) from Transduction
>laboritories.  Has anybody had experience with this antibody?  Is there a
>better one?  Does it require drastic retrieval methods?
>
>Any help or comments are appriciated!!!!
>
>Thanks,
>Kathy