Millipore Sigma Vibrant Logo
Attention: We have moved. EMD Millipore products are no longer available for purchase on emdmillipore.com.Learn More

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.
View Products on Sigmaaldrich.com
487664
Purchase on Sigma-Aldrich

Overview

Replacement Information

Products

Catalog NumberPackaging Qty/Pack
487664-500ML Plastic ampoule 500 ml
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
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
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
Global Trade Item Number
Catalog Number GTIN
487664-500ML 04055977200294

Documentation

p-Nitrophenyl Phosphate Substrate Buffer SDS

Title

Safety Data Sheet (SDS) 

p-Nitrophenyl Phosphate Substrate Buffer Certificates of Analysis

TitleLot Number
487664
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 Protect from light
+2°C to +8°C
Do Not Freeze Ok to freeze
Toxicity Standard Handling