Re: DAB film

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From:Katie B <>
To:Brandon Schanbacher <>, "''" <>
Date:Wed, 19 May 1999 10:35:11 -0700 (PDT)
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I too have a similar problem but with Vector Red leaving a red haze
across the entire slide for a new antibody I am using.  I also use the
Microprobe Probe on plus slides and almost want to say that my primary
is responsible for the haze (my negative control slides show nothing). 
I'm using a mouse monoclonal antibody with the Vector alk phos kit. 
The red haze doesn't interfere with detecting the positivitly labeled
cells however, so I've just kinda ignored it.  And doing the Levismole
(sp?) block step as recomended by Vector doesn't help.

Hmmm, as I write this, I'm wondering if I should do a pre-block with
normal mouse serum?  Something to try next staining session.


--- Brandon Schanbacher <> wrote:
> We have had some problems with DAB solution leaving a light brown
> haze
> across the entire slide during immunostaining.  If anyone has any
> suggestions on how to remedy this situation, I'd appreciate any
> input.
> The haze is most apparent when we use Upstate Biotechnologies' anti
> 3-nitrotyrosine antibody.  As far as our staining procedure, we use
> the
> Microprobe capillary gap system, which made us think it was a problem
> rinsing one of the antibody steps off of the slide.  We have
> increased
> the buffer rinses to no avail.  We don't get this film using other
> antibodies run at the same time, in the same DAB solution, so I doubt
> it's a problem with the DAB solution.  Has anyone else run into a
> similar situation?

Catherine "Katie" Bresee Bennett
Laboratory for Experimental Pathology
Department of Veterinary Pathology
Michigan State University

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