BUF044B is a high performance p-nitrophenol phosphate (pNPP) solution, recommended for use in ELISA as a substrate for alkaline phosphatase (AP). In the presence of AP, pNPP is hydolyzed rapidly to p-nitrophenol and inorganic phosphate to produce a yellow solution.
This ready to use solution has been carefully formulated to increase pNPP activity and stability. BUF044B may be used for both kinetic and endpoint tests.
Application
This product has been reported to work in the following applications. This information is derived from testing within our laboratories, peer-reviewed publications or personal communications from the originators. Please refer to references indicated for further information. For general protocol recommendations, please visitwww.abdserotec.com/protocols.
Application Name
Yes
No
Not Determined
Suggested Dilution
ELISA
Neat
Where this product has not been tested for use in a particular technique this does not necessarily exclude its use in such procedures. Suggested working dilutions are given as a guide only. It is recommended that the user titrates the product for use in their own system using appropriate negative/positive controls.
Storage
Store at +4oC. DO NOT FREEZE.
This product is photosensitive and should be protected from light.
Avoid exposure to light and heat
Store aliquots in bottles made of High Density Polyethylene (HDPE).
Shelf Life
12 months from date of despatch.
Instructions For Use
1. Remove the pNPP substrate container from the refrigerator, and allow it to reach room temperature prior to use.
2. Add 100ul of BUF044B per microtiter well. Development time is typically 15-30 minutes. Optimum time is determined by the user and will vary according to the procedure and the incubation temperature.
3. For kinetic tests, measure absorbance at 405nm. For endpoint tests, add 100ul 1.0M sodium hydroxide solution to each well and mix throughly prior to measuring absorbance at 405nm /650nm.
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