Vector's NovaRed

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From:Gayle Callis <uvsgc@msu.oscs.montana.edu>
To:histonet@Pathology.swmed.edu
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Date:Mon, 28 Jun 1999 15:59:40 -0600
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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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