[Histonet] RE: PBS or TBS?

[C.M. van der Loos]

Dear Joost,
As far as I know there is no difference in using PBS or TBS as washing buffer. Certainly not in terms of "killing" as you wrote. IHC is a very tolerant technique in this respect! 
The only situation to be careful with is when using alkaline phosphatase as marker enzyme. The enzymatic activity of this enzyme is heavily inhibited by phosphate ions in PBS. In this case you have to rinse your slides 3 times with a non-phosphate containing buffer (Tris-buffer for example) before starting the AP visualization reaction. 

Chris van der Loos
Dept. of Cardiovascular Pathology
Academical Medical Center
Amsterdam - The Netherlands

----- Original Message ----- 
>From  "Bruijntjes, J.P." <bruyntjes <@t> voeding.tno.nl> 
Date  Fri, 12 Dec 2003 15:55:18 +0100 
To  <histonet <@t> lists.utsouthwestern.edu> 
Subject  [Histonet] (no subject) 

Hi all
Can someone tell me, the type of buffer you use in immunocytochemistry
can it be killing for a successful staining? I mean it in the way that
an antibody will react when PBS is used instead of TBS, and the other
way around?

Joost Bruijntjes
TNO Nutrition and Food Research
PO Box 360
3700 AJ
Zeist
The Netherlands