487664
p-Nitrophenyl Phosphate Substrate Buffer
This p-Nitrophenyl Phosphate Substrate Buffer is validated for use in ELISA.
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| Pack Size | SKU | Availability | Price |
|---|---|---|---|
| 500 mL | Check Cart for Availability | CZK 4,990.00 |
About This Item
NACRES:
NA.77
UNSPSC Code:
41116158
Form:
liquid
Storage condition:
OK to freeze, protect from light
Quality Segment
sterility
non-sterile
form
liquid
manufacturer/tradename
Calbiochem®
storage condition
OK to freeze, protect from light
dilution
(ELISA )
color
clearlight yellow
shipped in
wet ice
storage temp.
2-8°C
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
Alkaline phosphatase substrate buffer
Product does not compete with ATP.
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.
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
Disclaimer
Toxicity: Standard Handling (A)
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This Item | |||
|---|---|---|---|
| form liquid | form crystalline solid | form solid | form - |
| Quality Level 100 | Quality Level 100 | Quality Level 100 | Quality Level 100 |
| manufacturer/tradename Calbiochem® | manufacturer/tradename Calbiochem® | manufacturer/tradename Calbiochem® | manufacturer/tradename Calbiochem® |
| storage temp. 2-8°C | storage temp. 2-8°C | storage temp. 2-8°C | storage temp. −20°C |
| storage condition OK to freeze, protect from light | storage condition OK to freeze, protect from light | storage condition OK to freeze | storage condition OK to freeze |
| color clearlight yellow | color light yellow | color off-white to pale yellow | color - |
signalword
Danger
hcodes
Hazard Classifications
Eye Dam. 1 - Repr. 2 - Skin Irrit. 2
Storage Class
12 - Non Combustible Liquids
wgk
WGK 2
flash_point_f
Not applicable
flash_point_c
Not applicable
Certificates of Analysis (COA)
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Global Trade Item Number
| SKU | GTIN |
|---|---|
| 487664-500ML | 04055977200294 |