[Histonet] RE: Buffer/Alkaline phosphatase stainng sytems

[C.M. van der Loos]

Dear Rena,
You are right about the fact that phosphate ions inhibit alkaline phosphatase activity. Long time ago we have tested this in practice and since that time we are dedicated TBS users. I have mentioned it in the Immunoenzyme Multiple Staining Methods handbook no.45 (Appendix E.1.2 AP, non-commercial visualization systems).
I hope this helps.

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

----- Original Message ----- 
>From  Rena Fail <RFail <@t> Charleston.net> 
Date  Mon, 09 Feb 2004 11:50:40 -0500 
To  histonet <@t> lists.utsouthwestern.edu 
Subject  [Histonet] FW: Buffer/Alkaline phosphatase stainng sytems 

We perform IHC on the DAKO Autostainer, at present we am using a
commercial concentrate of TRIS buffer with Tween . We are considering
making our buffers.
I have been told that phosphate buffers are not compatible with alkaline
phosphatase staining systems. Does anyone know of any references to
buffer/alkaline phophatase incompatibility?
Thank you
Rena Fail