p-nitropenyl phosphate (PNPP) substrate tabs and Buffer for ELISA
Description:
pNPP is a soluble substrate used in ELISA and for determination of alkaline and acid phosphatase in physiological fluids. For determination of acid phosphatase low pH buffer is required, not supplied.
Application:
ELISA
Presentation:
20 tablets
10X buffer (25 mls) (For alkaline phosphatase).
Aliquoting Instructions:
Dissolve each tablet in 5-10 ml of 1X buffer.
Staining Procedure:
ELISA PROCEDURE: Dissolve each tablet in 5-10 mls of 1X buffer. (dilute 10X buffer in distilled or deionized water). Add 5o microliters to each ELISA well and react for 30-60 mins. The reaction can be stopped with 50 microliters of 3 N NaOH and the yellow soluble color is read in spectrophotometer at 405 nm (ELISA reader)
Reactivity:
Reacts with alkaline phosphatase ( buffer supplied) and acid phosphatase (buffer pH about 5 not supplied).
Storage:
2-8 C
Technical Information:
p-nitrophenol (MW 139.1) is the hydrolysis product of PNPP and can be used as a std. to determine enzyme activity.
It can also be used to make standard curve for quantitation of enzyme.