Millipore Sigma Vibrant Logo
Atención: Nos hemos mudado. Los productos Merck Millipore ya no pueden adquirirse en MerckMillipore.comMás información

487663 p-Nitrophenyl Phosphate

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

p-Nitrophenyl Phosphate: Ficha de datos de seguridad (MSDS o SDS), certificado de análisis y de calidad (CoA y CoQ), expedientes, folletos y otros documentos disponibles.

Sinónimos: pNPP

487663
Purchase on Sigma-Aldrich

Descripción

Replacement Information

Products

Número de referenciaEmbalaje Cant./Env.
487663-10ML Frasco de vidrio 10 ml
Description
OverviewExcellent 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 Number487663
Brand Family Calbiochem®
SynonymspNPP
References
Product Information
ATP CompetitiveN
FormClear to slightly yellow tinged liquid
FormulationSupplied as a 50X concentrate.
PreservativeNone
ReversibleN
Quality LevelMQ100
Applications
Application NotesELISA
Application CommentsFunctionality 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 dH2O 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.
Biological Information
Primary TargetSubstrate for alkaline phosphatase-based ELISA assays
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 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 InstructionsDiscard if solution turns bright yellow or becomes turbid.
Packaging Information
Transport Information
Supplemental Information
Specifications
Global Trade Item Number
Número de referencia GTIN
487663-10ML 04055977200287

Documentation

p-Nitrophenyl Phosphate Ficha datos de seguridad (MSDS)

Título

Ficha técnica de seguridad del material (MSDS) 

p-Nitrophenyl Phosphate Certificados de análisis

CargoNúmero de lote
487663

Folleto

Cargo
Protein Phosphatases Technical Bulletin
Ficha técnica

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
SynonymspNPP
ApplicationELISA
Descriptionp-Nitrophenyl phosphate supplied as a 50X concentrate (0.19 mol/l). For use with p-NPP Substrate Buffer, Cat. No. 487664.
Backgroundp-Nitrophenyl phosphate is the most common substrate for alkaline phosphatase-based ELISA assays. Hydrolysis of the phosphate ester liberates p-nitrophenol, a soluble, yellow chromogen that can be measured spectrophotometrically at 405 nm (ε = 1.83 x 104 M-1cm-1). As the reaction is linear, kinetic assays can be performed. Endpoint determinations can also be performed by stopping the reaction with strong base and measuring the absorbance at 405 nm. This p-NPP preparation is a stable, convenient, liquid concentrate in a proprietary solvent.
FormClear to slightly yellow tinged liquid
FormulationSupplied as a 50X concentrate.
PreservativeNone
CommentsFunctionality 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 dH2O 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.
Storage Protect from light
+2°C to +8°C
Do Not Freeze Ok to freeze
Special InstructionsDiscard if solution turns bright yellow or becomes turbid.
Toxicity Standard Handling