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Merck - 487663-10ML - P-NITROPHENYL PHOSPHAT 10ML (Each)

  • MFR SKU: 487663-10ML
  • ITEM #: 3441936
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ITEM #: 3441936
This p-Nitrophenyl Phosphate is validated for use in ELISA.
E x cellent substrate for alkaline phosphatase-based ELISA assays. Produces yellow soluble end product that can be read at 405 nm to 410 nm. Both kinetic and endpoint determinations can be performed. Supplied as ready-to-use.
Supplied as a 50X concentrate.
p-Nitrophenyl Phosphate, 10mL
This listing is for Each

Merck - 487663-10ML - P-NITROPHENYL PHOSPHAT 10ML (Each)

  • Item #: 3441936
  • MFR SKU: 487663-10ML
Technical Specs
  • Overview Excellent substrate for alkaline phosphatase-based ELISA assays. Produces yellow soluble end product that can be read at 405 nm to 410 nm. Both kinetic and endpoint determinations can be performed. Supplied as ready-to-use. A ready-to-use diluent is available separately (Cat. No. 487664 ).
  • Catalogue Number 487663
  • Brand Family Calbiochem®
  • Synonyms p NPP
  • ATP Competitive N
  • Form Clear to slightly yellow tinged liquid
  • Formulation Supplied as a 50X concentrate.
  • Preservative None
  • Reversible N
  • Quality Level MQ100
  • Application Notes ELISA
  • Application Comments Functionality check performed on the final product in ELISA.
  • Suggested procedure for use of p -NPP in alkaline phosphatase-based ELISAs: 1. Prepare 1X p -NPP Substrate Solution: To 39 parts dH 2 O add 1 part 50X p -NPP Concentrate and 10 parts p -NPP Substrate Buffer (Cat. No. 487664). Mix well and store protected from light. This diluted reagent is suitable for up to 10 h after preparation.
  • Note: Cat. No. 487664 is an AMP (2-amino-2-methyl-1-propanol)-based buffer. Other buffers, e.g. diethanolamine (DEA)-based and 2-amino-2-methyl-1, 3-propanediol (AMPD)-based, may also be employed.
  • 2. Complete all required incubations with antibodies and alkaline phosphatase-labeled probes.
  • 3. Wash plate wells at least 4 times with Tris-buffered saline (TBS) containing 0.1% Tween ® -20 detergent. DO NOT USE PHOSPHATE BUFFERS; inorganic phosphate is a potent inhibitor of alkaline phosphatase.
  • 4. After the final wash, shake and blot all residual buffer from the wells.
  • 5. Add 100 µl of 1X p -NPP Substrate Solution (prepared in Step 2) to each well and incubate for 15-60 min. (If a kinetic study is desired, readings at 405 nm may be taken at intermediate intervals).
  • [Note: Optimal incubation times may vary depending on the amount of enzyme present. If color develops too quickly, dilution of the probe, antibody, or alkaline phosphatase-labeled reagent may be required.]
  • 6. Stop the reaction by adding 100 µl 1 N NaOH.
  • 7. Read absorbance at 405 nm.
  • Note: Variations in time, reagent volumes, and temperature may require further standardization by the user.
  • Primary Target Substrate for alkaline phosphatase-based ELISA assays
  • Cell permeable N
  • Ship Code Ambient Temperature Only
  • Toxicity Standard Handling
  • Storage +2° C to +8° C
  • Protect from Light Protect from light
  • Do not freeze Ok to freeze
  • Special Instructions Discard if solution turns bright yellow or becomes turbid.
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