Okadaic Acid?

Jonathan Mazer szmazer at chip.ucdavis.edu
Tue Sep 12 17:09:51 EST 1995


Hi,

   I am getting ready to label (yeast) cells with 32P to see if my 
(membrane) protein is in vivo phosphorylated. We think it may be 
phosphorylated, but not by the cyclin kinases (possibly PK-A). My 
questions is : 

1.) Will okadaic acid make any difference in inhibiting phosphatase 
    activity even if the active kinase is not one of the cyclin 
    kinases? 

2.) Do the phosphatases which are specific for cyclin kinase substrates 
    also dephosphorylated other phophoproteins once a whole cell extract  
    is created? 

3.) (Is okadaic acid only specific for the substrates of the cyclin kinases?)


    I plan to use NaF as the phosphatase inhibitor after the cells are 
lysed, but I was just wondering if I could save some money by not using 
okadaic acid if it isn't really necessary. (OA is extremely expensive! 
100 ug = $350!!)

Thanks for any help.

   -Jonathan Mazer (jsmazer at ucdavis.edu)
    



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