Description:Para-nitrophenyl phosphate (pNPP) is a chromogenic substrate for most phosphatases such as alkaline phosphatases, acid phosphatases, protein tyrosine phosphatases and serine/threonine phosphatases. The reaction yields para-nitrophenol, which becomes an intense yellow soluble product under alkaline conditions and can be conveniently measured at 405 nm on a spectrophotometer. phosphatase p-Nitrophenyl phosphate p-nitrophenol + phosphateThis homogeneous mix-and-measure assay involves simply adding a single reagent to the phosphatase and measuring the product formation using any absorbance reader. The assay can be conveniently performed in cuvettes, tubes or multi-well plates at either room temperature or 37°C. In addition, the reagents are compatible with ELISA assays in which alkaline phosphatase conjugated secondary antibody is used. This kit is well adapted for a number of applications. For example, it can be utilized for direct characterization of enzyme activity and for assay condition optimization. It can also be applied for research diagnosis of diseases that are associated with increased levels of alkaline phosphatase. Typical diseases include liver disease, bone disease, Hodgkin's disease, congestive heart failure, Fanconi's syndrome, hyperparathyroidism, intestinal disease and abdominal bacterial infections. Moreover, it can be used to characterize phosphatase inhibitors through high-throughput screening.The kit reagents have been optimized for long shelf life and maximum reproducibility. The reagents are compatible with all liquid handling systems and bulk reagents are available for high-throughput screening of phosphatase inhibitors.Key Features:High sensitivity and wide linear range. The detection limit is generally 3 ng phosphatase or below.Homogeneous and simple procedure. No wash or reagent transfer steps are involved. The assay can be completed within 30 minutes.Robust and amenable to HTS. All reagents are compatible with high-throughput liquid handling instruments.Applications:Enzyme Activity Assay and Quality Control for phosphatase production. Characterization of Kinetics of phosphatase reaction.Drug Discovery: high-throughput screen for phosphatase inhibitors.Kit Contents:Size (assays)ReagentAssay BufferStop Solution500solid25ml25ml1,000solid50ml50ml2,400solid120ml120mlStorage conditions. The Reagent should be stored in the amber tube at -20°C. The Assay Buffer and Stop Solution are stable at 4°C and room temperature, respectively. Shelf life: 12 months.Precautions: reagents are for research use only. Normal precautions for laboratory reagents should be exercised while using the reagents. Please refer to Material Safety Data Sheet for detailed information.
