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487664 p-Nitrophenyl Phosphate Substrate Buffer

This p-Nitrophenyl Phosphate Substrate Buffer is validated for use in ELISA.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.
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487664
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Overview

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487664-500ML
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      Description
      OverviewAlkaline phosphatase substrate buffer supplied as ready-to-use. A diluent for Cat. No. 487663.
      Catalogue Number487664
      Brand Family Calbiochem®
      Product Information
      ATP CompetitiveN
      FormClear to light yellow liquid.
      ReversibleN
      Quality LevelMQ100
      Applications
      Application NotesELISA (see comments)
      Application CommentsProduct tested for stability at 4°C, 18-26°C, and 37°C. Product performance is tested on the final product by ELISA.

      Suggested procedure for use of p-NPP in alkaline phosphatase-based ELISAs:

      1. To 39 parts distilled water add 1 part 50X p-NPP Concentrate (Cat. No. 487663) and 10 parts p-NPP Substrate Buffer. Mix well and store protected from light. This diluted reagent is suitable for up to 10 h after preparation.
      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 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 phophatase-labeled reagent may be required.]
      6. Stop the reaction by adding 100 µl of 1 N NaOH.
      7. Read absorbance at 405 nm.

      Note: Variations in time, reagent volumes, and temperature may require further standardization by the user.
      Biological Information
      Primary TargetAlkaline phosphatase substrate buffer
      Physicochemical Information
      Cell permeableN
      Storage and Shipping Information
      Ship Code Blue Ice Only
      Toxicity Standard Handling
      Storage +2°C to +8°C
      Protect from Light Protect from light
      Do not freeze Ok to freeze
      Global Trade Item Number
      Catalogue Number GTIN
      487664-500ML 04055977200294