487664 | p-Nitrophenyl Phosphate Substrate Buffer

487664
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      Overview

      Replacement Information

      Key Specifications Table

      Pricing & Availability

      Catalog NumberAvailability Packaging Qty/Pack Price Quantity
      487664-500ML
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      Stocked 
      Discontinued
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      Available
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          Plastic ampoule 500 ml
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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®
          References
          Product Information
          ATP CompetitiveN
          FormClear to light yellow liquid.
          ReversibleN
          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
          Dimensions
          Materials Information
          Toxicological Information
          Safety Information according to GHS
          Safety Information
          Product Usage Statements
          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
          Packaging Information
          Transport Information
          Supplemental Information
          Specifications

          Documentation

          SDS

          Title

          Safety Data Sheet (SDS) 

          Certificates of Analysis

          TitleLot Number
          487664

          Brochure

          Title
          Enzyme-Linked Detection Systems Technical Bulletin
          Data Sheet

          Note that this data sheet is not lot-specific and is representative of the current specifications for this product. Please consult the vial label and the certificate of analysis for information on specific lots. Also note that shipping conditions may differ from storage conditions.

          Revision20-November-2009 JSW
          ApplicationELISA (see comments)
          DescriptionAlkaline phosphatase substrate buffer supplied as ready-to-use. A diluent for Cat. No. 487663.
          FormClear to light yellow liquid.
          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.
          Storage +2°C to +8°C
          Protect from light
          Do Not Freeze Ok to freeze
          Toxicity Standard Handling