method for measurement of alkaline phosphatase ...

[Dr Engelbert Buxbaum]

Am 08.10.2008, 03:50 Uhr, schrieb Ebrahim Sotoudeh  
<ebrahimsotoudeh from gmail.com>:

> I want to assay alkaline phosphatase activity. and I need method for *
> measurement* of *alkaline phosphatase* activity .

The cheapest way to do this is to follow the hydrolysis of  
para-nitrophenylphosphate (pNPP) (colourless) to nitrophenol (yellow in  
alkaline solutions) photometrically at 405 nm. Acid phosphatase is  
measured in citrate buffer pH 5.6, alkaline phosphatase at pH 8-11 (you  
need to find out the optimum for your enzyme, check BRENDA for published  
data). If sample size are limited you can use methylumbelliferyl phosphate  
instead, which gives a fluorescent product.