endogenous alkaline phosphatase activity

lgd450 at nwu.edu lgd450 at nwu.edu
Mon Oct 14 16:52:59 EST 1996


Does anyone know of a good method for blocking endogenous alkaline
phosphatase activity in immunohistochemistry?  I am using frozen sections
(40 microns thick) of human tissue that are in some procedures mounted
prior to staining and sometimes free floated.  I seem to be getting a lot
of background (glia and blood vessles mostly).  When I do peroxidase
reactions with the same antibodies, there is almost no background.

The company providing the AP-kit and substrate suggest putting Levamisole
in the buffer used to make the substrate.  How does this work?  I am
worried about altering otherwise nice staining by adding new compounds!

Is it possible to treat the tissue with phosphatase inhibitors and still
get reliable staining?

Any help is much appreciated!

Thank you in advance,

Angie

-- 
The attempt to develop a sense of humor and to see things in a humorous light is some kind of a trick learned while mastering the art of living.

----Viktor E. Frankl   in "Man's Search for Meaning"



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