endogenous alkaline phosphatase activity

Kathie Berghorn kab35 at cornell.edu
Tue Oct 15 14:31:28 EST 1996


lgd450 at nwu.edu wrote:
> 
> 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"


Angie,
	We have used levamisole and alkaline phosphatase consistently 
with free floating fixed tissue sections with great success.  Right now 
we are using Sigma's Fast Red as the chromogen and can do triple staining 
successfully-i.e. no interference from anything in our solutions, inc, 
Levamisole.  Kathie



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