Re: iNOS

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From:Jamie Erickson <>,
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            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 <> 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=20
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!!!!


Kathy Walters                                // 
Central Microscopy Research Facility        / /
85 EMRB                                    / /\
University of Iowa                        / /\ \
Iowa City, Iowa 52242                    / /  \ \
Phone #:  (319) 335-8142                / /    \ \
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Jamie Erickson
Associate Scientist
Genetics Institute
1 Burtt Rd.
Andover, MA   01810
work : (978) 247-1348
FAX  : (978) 247-1333

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