Re: detergents in antibody diuents

From:Connie McManus <conmac@cc.usu.edu>

I use tween 20 with TRIS buffer for all my rinses (IHC).  I've never had
a problem with sections coming off, however, I find that the surface
tension on the slide is nill and unless I do a plain TRIS rinse, all the
reagent runs off the slide. 

I use the tween 20 because i was told that it helps remove the excess
reagent so you get cleaner rinsing between reagents.  At one time, I
used only plain TRIS buffer to rinse in, however, there was some
nonspecific background staining that was hard to interpret.  Doing the
same number of rinses as before, the nonspecific background staining is
gone since using the tween 20. 

Connie McManus

Neuropathology wrote:
> 
>  We have used Tween 20 in our antibody diluent at a concentration  of 0=2E05%
> without any problems on both paraffin and frozen sections.
> 
> Bob Quilty
> Dept of Neuropathology
> Frenchay Hospital
> Bristol  UK
> 
> ----- Original Message -----
> From: Ron Salisbury <ronald2@uakron.edu>
> To: Histonet <histonet@pathology.swmed.edu>
> Sent: Tuesday, May 01, 2001 7:06 PM
> Subject: detergents in antibody diuents
> 
> > I have seen various concentrations of detergents (Triton X, Tween 20
> > etc.) used in primary antibody diluents and wondered if there were any
> > guidelines for choosing a particular concentration. Does it depend on
> > whether paraffin or frozen sections are used or is it more dependent
> > upon the type and concentration of fixative? Also, is there an effect of
> > detergent concentration on the ability of sections to adhere to slides?
> >
> >

-- 
ÐÏࡱá



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