p-Nitrophenylphosphate is hydrolyzed to form p-nitrophenol and phosphate in the presence of magnesium ions and alkaline phosphatase in a 2-amino-2-methyl-1-propanol [AMP] buffer.
alk
phos
p-nitrophenylphosphate -> p-nitrophenol + phosphate
Mg+2 (yellow)
The rate of appearance of p-nitrophenol measured at 405 nm is directly proportional to the alkaline phosphatase activity present in the original sample.
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