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PNPP Substrates


An alkaline phosphatase substrate for ELISA applications.

Pierce PNPP Substrates

Thermo Scientific Pierce PNPP Substrates are powders, tablets, solutions and complete kits of p-nitrophenyl phosphate, a colorimetric, soluble substrate of alkaline phosphatase for use in ELISA applications.

PNPP (p-Nitrophenyl Phosphate, Disodium Salt) is a widely used substrate for detecting alkaline phosphatase in ELISA applications. When alkaline phosphatase and PNPP are reacted, a yellow water-soluble reaction product is formed. This reaction product absorbs light at 405nm. The substrate is offered in several formats.

Highlights:

  • One-component alkaline phosphatase (AP) substrate
  • Absorbance maximum of 405nm
  • Develops a yellow hue in solution upon reaction with AP
  • Safer to use — contains a non-mercury, non-azide preservative

Product Details:

PNPP is available either as a crystalline powder or 5mg tablets. Also available is the Phosphatase Substrate Kit, which contains PNPP tablets and a Diethanolamine Buffer to yield more than one liter of substrate. The Diethanolamine Substrate Buffer (Part No. 34064) is also provided individually as a 5X concentrate. The Pierce formulation has an optimal and consistent pH and it is stable even at a 1X concentration.

1-Step PNPP is also available. Until now, the method for preparing PNPP solutions required dissolving the powder or tablets in buffer and then diluting the solution to the desired concentration. Substrate instability made it necessary to prepare it on a daily basis. Pierce 1-Step PNPP circumvents this time-consuming and variable introducing step by providing a single component PNPP substrate. This substrate is stable for 12 months at 2°C-8°C and can be stopped with conventional methods.

References:

  1. Snyder, S.L., et al. (1972). Biochim. Biophys. Acta 258, 178-187.
  2. Bosque, P.J., et al. (2002). Proc. Natl. Acad. Sci. USA 99, 3812-3817.
  3. Jan, J.-T., et al. (2000). J. Virol. 74, 8680-8691.

Related Resources:

Assay development and ELISA technical guide
ELISA substrate selection guide


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
34045 PNPP
Formulation: p-Nitrophenyl phosphate disodium salt; crystalline powder
Sufficient For: 25L of typical 1X (1mg/mL) substrate solution
25g Product Instructions for product #34045 PNPP MSDS for product #34045 PNPP Certificate of Analysis for product #34045 PNPP $131.00

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34045B PNPP
Formulation: p-Nitrophenyl phosphate disodium salt; crystalline powder
Custom MSDS for product #34045B PNPP Certificate of Analysis for product #34045B PNPP
34047 PNPP Tablets
Formulation: Tablets each containing 5mg of PNPP with inert filler as the balance
Sufficient For: 5mL of typical 1X substrate solution per tablet (525mL total)
105 tablets Product Instructions for product #34047 PNPP Tablets MSDS for product #34047 PNPP Tablets Certificate of Analysis for product #34047 PNPP Tablets $133.00

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37620 PNPP Substrate Kit
Formulation: Two-component kit
Sufficient For: 5mL of typical 1X substrate solution per tablet (525mL total; approx. 5250 wells at 100μL per well)

Kit Contents:
Diethanolamine Buffer, 225mL
PNPP Tablets, 105 tablets (5mg/tablet)
105-tablet kit Product Instructions for product #37620 PNPP Substrate Kit MSDS for product #37620 PNPP Substrate Kit Certificate of Analysis for product #37620 PNPP Substrate Kit $173.00

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37621 1-Step PNPP Solution
Formulation: Ready-to-use (1X) proprietary buffered solution of p-Nitrophenyl phosphate disodium salt
Sufficient For: 1000 wells at 100μL per well
100mL Product Instructions for product #37621 1-Step PNPP Solution MSDS for product #37621 1-Step PNPP Solution Certificate of Analysis for product #37621 1-Step PNPP Solution $103.00

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37621B 1-Step PNPP Solution
Formulation: Ready-to-use (1X) proprietary buffered solution of p-Nitrophenyl phosphate disodium salt
Custom MSDS for product #37621B 1-Step PNPP Solution Certificate of Analysis for product #37621B 1-Step PNPP Solution

Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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