Re: Vector's NovaRed

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From:Michelle M Sutorik <istsmmmc@umich.edu>
To:Gayle Callis <uvsgc@msu.oscs.montana.edu>
Reply-To:
Date:Mon, 28 Jun 1999 21:55:06 -0400 (EDT)
Content-Type:TEXT/PLAIN; charset=US-ASCII

Hi Gayle, 
I am writing rom home and the server is slow so forgive me if i dont
correct the mistakes.  first fo all, you are right , the nova red is not
compatible with an aqueous mount.  I started using the nova red a few
weeks ago and noticed the same things.  Here is the sololution to the
problem.  First of all, start with using histological grade ethanol to
dehydrate.   Start at 95% then 100%x2 each 1min, then   3 times in  xylene
for 3min each.  Oh by the way use  deionized h20 for the rinse.  Dont need
to have the water runing when you rinse.  


I love it too..  It is incredibly sensive and i am amazed how muchg my
antibody titer has decreased.  let me know if i can be of further
assistance.  Have a great day./

Michelle M. Sutorik
oral pathology/oncology


 On Mon, 28 Jun 1999, Gayle Callis wrote:

> This question also went to Vector and in the process, answered itself
> as I type.  This new DAB (red color) substitute is wonderful, and 
> worked beautifully with acetone fixed mouse CD markers CD4, 8 , Mac1,
> and B220 but after rinsing in tap water, counterstaining and
> subsequent mounting in aqueous mounting media (Shandon Immunomount)
> 
> totally faded within hours.  Bummer of bummers!  I guess aqueous mounting
> media is not acceptable with this chromogen, you need to dehydrate and
> use a regular mounting media.  Also, directions say to rinse sections
> for 5 min in water.  Question here:  running tap water?  distilled water?
> is this the source of fading also?  I know you need to avoid recycled
> alcohols with this chromogen.  It also says mounting with non aqueous mounting
> media acceptable, no mention of aqueous, but did not say no on the latter. 
> 
> One good thing, NovaRed is so sensitive, that antibody dilutions used with
> AEC chromogen were too concentrated, and a mouse CD4 at 1:1000 was still
> too concentrated.  Impressed with the sensitivity, but not the fading.
> 
> Has anyone else had this happen?  or tried it with aq mounting media?
> 
> So many questions, so little time!  
> 
> Gayle Callis, 
> 




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