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
Code UNSPSC :
41116158
Produits recommandés
Niveau de qualité
Stérilité
non-sterile
Forme
liquid
Fabricant/nom de marque
Calbiochem®
Conditions de stockage
OK to freeze
protect from light
Couleur
clearlight yellow
Conditions d'expédition
wet ice
Température de stockage
2-8°C
Description générale
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.
Application
ELISA (see comments)
Actions biochimiques/physiologiques
Primary Target
Alkaline phosphatase substrate buffer
Alkaline phosphatase substrate buffer
Product does not compete with ATP.
Avertissement
Toxicity: Standard Handling (A)
Autres remarques
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.
Informations légales
CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany
TWEEN is a registered trademark of Croda International PLC
Mention d'avertissement
Danger
Mentions de danger
Conseils de prudence
Classification des risques
Eye Dam. 1 - Repr. 2 - Skin Irrit. 2
Code de la classe de stockage
12 - Non Combustible Liquids
Classe de danger pour l'eau (WGK)
WGK 2
Point d'éclair (°F)
Not applicable
Point d'éclair (°C)
Not applicable
Certificats d'analyse (COA)
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