p-Nitrophenyl Phosphate (pNPP), 50X Concentrate, 0.19M

Excellent substrate for phosphatase based ELISA assays. It produces a soluble end product which is yellow in color and read at 405nm to 410nm. Both kinetic and end point determinations can be performed. Supplied as a 50X concentrate. Dual component system.\n\npNPP is the most conventional reagent employed for estimating alkaline phosphatase (AP) activity. It is usually provided in powder or tablet form and dissolved in buffer prior to use. Working concentrations range from 3.5-4.0mM when used for ELISA assays.\n\nAfter completion of analyte binding to a solid phase and reaction with an AP labeled probe, diluted pNPP Solution is added. Hydrolysis of the phosphate ester liberates p-nitrophenol producing a yellow chromogen with an extinction coefficient of 1.83 x 10e4 mol-1cm-1 at 405nm.\n\nThe reaction is linear allowing kinetic assays. Endpoint analysis can be performed by stopping the reaction with strong base and reading the final color at 405nm. Transphorylating agents such as diethanolamine (DEA), 2-amino-2-methyl-1-propanol (AMP) or on some occasions 2-amino-2-methyl-1,3-propanediol (AMPD) are the prevalent buffers.\n\nFor maximum activity: dilute in N2600-15 \n\nColor and Clarity:\nColorless to slight yellow\n\nStability (4 degrees C):\nTo pass specifications\n\nAbsorbance:\nTo pass specifications\n\nELISA Performance:\nTo pass specifications\n\nAlso Available:\nN2600: p-Nitrophenyl Phosphate (pNPP)\nN2600-10: p-Nitrophenyl Phosphate (pNPP), 50X Concentrate, 0.19M/Liter\nN2600-15: p-Nitrophenyl Phosphate (pNPP) Buffer, DEA

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