Re: iNOS
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From: | Jamie Erickson <JErickson@genetics.com> |
To: | kwalters@blue.weeg.uiowa.edu, histonet@pathology.swmed.edu |
Reply-To: | |
Content-Type: | text/plain; charset=US-ASCII |
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=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!!!!
Thanks,
Kathy
Kathy Walters //
Central Microscopy Research Facility / /
85 EMRB / /\
University of Iowa / /\ \
Iowa City, Iowa 52242 / / \ \
Phone #: (319) 335-8142 / / \ \
Fax #: (319) 335-6710 ______ ((0))
email: Katherine-Walters@uiowa.edu |__| / /
|| / /
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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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