Julie,

Try popping the coverslips, running them back to water and then placing them
in a 10% Cupric Chloride (& DI water) solution for ten minutes.  This may
enhance your DAB staining.

Glen Dawson

-----Original Message-----
From: rueggp [mailto:rueggp@earthlink.net]
Sent: Thursday, July 11, 2002 3:12 PM
To: Julie Clark; HistoNet Server
Subject: Re: faded DAB


Julie,
I have not had any success restaining sections, in my experience they either
get staining all over or none at all, but if these are precious sections I
suppose I would seek the experts advise and give it a try.  Perhaps before
restaining all together you could try some of the DAB enhancment methods if
they are already staining specifically just very light (faded).
Patsy

Julie Clark wrote:

> Hi Patsy,
> I know they are DAB because it is the grant protocol, that is why I can't
change chromgens.  It seems like I have been using DAB+ since 2000 (when the
slides originated), but it doesn't mean they were.  I have been doing immuno
for almost 13 years and have never seen anything like this.  Would you try
re-staining or do you think you would just get horrible background?
>
> Thanks,
> Julie
> >>> rueggp  07/10/02 11:21AM >>>
> Julie,
> Are you sure the orginal slides were DAB stained and are now faded?  I
have no experience with DAB fading, I have seen other chromogens fad such as
AEC.  Another
> thought is that with improved enhanced IHC detection systems maybe we are
just not used to looking at the lighter staining we used to get.
> Patsy
>
> Julie Clark wrote:
>
> > I have been asked to review some two year-old immuno slides.  Most of
the slides are very faded to completely faded.  I have no idea what brand
DAB they were using.
> >
> > Has anyone tried to re-stain previously DAB stained immuno slides?  I
have to use DAB, I cannot switch chromogens.
> >
> > Is there some way to remove the faded DAB?
> >
> > Thanks,
> >
> > Julie Clark
> > Technical Director
> > Breast Cancer Prevention Center Research Laboratory
> > Room 3021 Lied
> > Univ. of Kansas Med. Center
> > Kansas City, KS
> > Phone: Office: 913-588-3963
> >             Lab:     913-588-3917
> > FAX: 913-588-3821
> > Page: 913-917-6052