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Can be alkaline phosphatise inhibit by PBS?


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#1 asias

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Posted 15 January 2010 - 03:59 AM

Hi everyone

I work with phosphor-specific antibodies as a control a use alkaline phosphatase. Phosphatase work well when I use them directly on the blot or cell lisate but do not work when I use then in IP. The mine different between IP and the cell lisates is that IP is washed with PBS and as a results complex protein-antibodies-proteinA is in PBS.

Do you know that PBS can inhibit alkaline phosphatise? It is possible?

Edited by asias, 15 January 2010 - 04:01 AM.


#2 mdfenko

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Posted 15 January 2010 - 08:06 AM

yes. free phosphate will inhibit ap.
talent does what it can
genius does what it must
i do what i get paid to do

#3 bob1

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Posted 17 January 2010 - 03:12 PM

Use a tris- buffered wash instead of PBS.




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