487664
p-Nitrophenyl Phosphate Substrate Buffer
This p-Nitrophenyl Phosphate Substrate Buffer is validated for use in ELISA.
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About This Item
Codice UNSPSC:
41116158
Livello qualitativo
Sterilità
non-sterile
Forma fisica
liquid
Produttore/marchio commerciale
Calbiochem®
Condizioni di stoccaggio
OK to freeze
protect from light
Colore
clearlight yellow
Condizioni di spedizione
wet ice
Temperatura di conservazione
2-8°C
Descrizione generale
Alkaline phosphatase substrate buffer supplied as ready-to-use. A diluent for Cat. No. 487663.
This p-Nitrophenyl Phosphate Substrate Buffer is validated for use in ELISA.
Applicazioni
ELISA (see comments)
Azioni biochim/fisiol
Primary Target
Alkaline phosphatase substrate buffer
Alkaline phosphatase substrate buffer
Product does not compete with ATP.
Attenzione
Toxicity: Standard Handling (A)
Altre note
Product 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.
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.
Note legali
CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany
TWEEN is a registered trademark of Croda International PLC
Avvertenze
Danger
Indicazioni di pericolo
Consigli di prudenza
Classi di pericolo
Eye Dam. 1 - Repr. 2 - Skin Irrit. 2
Codice della classe di stoccaggio
12 - Non Combustible Liquids
Classe di pericolosità dell'acqua (WGK)
WGK 2
Punto d’infiammabilità (°F)
Not applicable
Punto d’infiammabilità (°C)
Not applicable
Certificati d'analisi (COA)
Cerca il Certificati d'analisi (COA) digitando il numero di lotto/batch corrispondente. I numeri di lotto o di batch sono stampati sull'etichetta dei prodotti dopo la parola ‘Lotto’ o ‘Batch’.
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