[Histonet] RE: NaCl in antibody diluent

From:"C.M. van der Loos"


Jennifer and Rene,

As Rene said the NaCl is part of the PBS. The 0.9% concentra= tion is equal to 0.15 M. I have heard (but never tested) of somebody = who raised the NaCl concentration to 0.5 M for reducing non-specific ba ckground staining. He used this high salt PBS not only for diluting the= primaries but also as washing buffer. Take care this high salt concent= ration only works for FFPE sections. It will certainly destroy= your acetone-fixed cryo's!


Chris van der Loos, PhD
Dept. of Pathology
Acade= mic Medical Center M2-230
Meibergdreef 9
NL-1105 AZ Amsterd= am
The Netherlands


Date: Tue, 27 Nov 2007 09:44:33 -0800 (PST)
From:= Rene J Buesa <rjbuesa@yahoo.com>
Subject: Re: [Histonet] NaCl in antibody diluent
To: "Harvey, Jennif= er Lynn" <jennifer.harvey@Vanderbilt.Edu>,

Jennifer:=   The Ab diluent is made of 1% bovine serum albumen in PBS wit= h ).05% Tween 20 and 0.1% sodium azide.'NaCl is part of the P= BS but I have never used or kbown anybody using additional NaCl
&= nbsp; René J.

"Harvey, Jennifer Lynn" < jennifer.harvey@Vanderbilt.Edu> wrote:
  H= istonet,

Has anyone tried NaCl in there diluent buffer f= or IHC, if so what
concentration. I read about it in DAKO's immunostaining methods booklet
but they don't state the concentr= ation. This is my last chance with my
background problem. I ha= ve a sticky collagen issue I cant get rid of.

Thanks!<= BR>

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