General description
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)
Biochem/physiol Actions
Primary Target
Alkaline phosphatase substrate buffer
Product does not compete with ATP.
Warning
Toxicity: Standard Handling (A)
Other Notes
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.
Legal Information
CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany
TWEEN is a registered trademark of Croda International PLC
Quality Level: 100. sterility: non-sterile. form: liquid. manufacturer/tradename: Calbiochem®. . storage condition: OK to freeze, protect from light. color: clearlight yellow. shipped in: wet ice. storage temp.: 2-8°C. Pictograms: GHS05,GHS08. Signal Word: Danger. Hazard Statements: H315 - H318 - H361fd. Precautionary Statements: P202 - P264 - P280 - P302 + P352 - P305 + P351 + P338 - P308 + P313. Hazard Classifications: Eye Dam. 1 - Repr. 2 - Skin Irrit. 2. Storage Class Code: 12 - Non Combustible Liquids. WGK: WGK 1. Flash Point(F): Not applicable. Flash Point(C): Not applicable.Shipping Information:
Dry Ice Surcharge & Ice Pack Shipments: $40
More Information: https://cenmed.com/shipping-returns
- UPC:
- 51372118
- Condition:
- New
- Availability:
- 3-5 Days
- Weight:
- 1.00 Ounces
- HazmatClass:
- No
- MPN:
- 487664-500ML
- Temperature Control Device:
- Yes
