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| Product Name | >=15 U/mg Purine Nucleoside Phosphorylase for 505 nm Coupled Assays |
| Catalog No. | IEC-HMM-0246 |
| Description | Alta DiagnoTech purine nucleoside phosphorylase is a pale-yellowish white amorphous enzyme powder for coupled biochemical reagent systems. |
| Intended Use | Enzyme raw material for adenosine deaminase and 5'-nucleotidase biochemical assay reagents. |
| Detection Method | Coupled kinetic colorimetric assay at 505 nm |
| Performance Range / Specifications | Specific activity >=15.0 U/mg; recommended assay sample range 200-300 U/L. |
| Reaction Conditions / Protocol | Prepare Reagent 1 in 800 mL water with 13.6 g potassium dihydrogen phosphate and 0.744 g EDTA disodium salt, adjust to pH 7.50 at 25 degrees C with KOH, then add 0.14 g sodium cholate, 0.24 g 4-aminoantipyrine, 4 kU xanthine oxidase and 1 g sodium azide; bring to 1 L and confirm pH 7.50 +/- 0.05. Prepare Reagent 2 in 800 mL water with 7.8 g sodium dihydrogen phosphate dihydrate and 11.4 g dipotassium hydrogen phosphate, adjust to pH 7.7, then add 3.22 g inosine, 2.8 g phenol, 1 g sodium azide and 10 kU peroxidase; bring to 1 L and adjust to pH 7.70 +/- 0.05. Combine 0.750 mL Reagent 1 and 0.150 mL Reagent 2, preincubate for 300 seconds at 37 degrees C, add 0.015 mL sample and measure the 1-minute absorbance-change rate at 505 nm. Activity in U/L equals Delta A/min x 4,586 x dilution factor. |
| Storage Conditions | Store at -20 degrees C |
| Shelf Life | At least 1 year at -20 degrees C |
| Product Form | Pale-yellowish white amorphous powder |
| Activity / Unit Definition | One unit is the amount of enzyme that produces 1 umol hydrogen peroxide per minute under the stated reaction conditions. |
| Molecular Weight | Approximately 29 kDa |
| pH Range / Optimal pH | Stable at pH 5.0-10.0; optimum pH 6.0 |
| Recommended Buffer System | Sample diluent: dissolve 13.6 g potassium dihydrogen phosphate in 800 mL water, adjust to pH 7.50 at 25 degrees C with KOH, add 0.744 g EDTA disodium salt, 0.14 g sodium cholate and 1 g sodium azide, then bring to 1 L. Allow prepared enzyme sample solution to stabilize for 1.5 hours at room temperature and complete the assay within 2 hours. |
| Appearance | Pale-yellowish white amorphous powder |
| Alias | Purine Nucleoside Phosphorylase |
| Product Type | Purine nucleoside phosphorylase enzyme raw material |
| Function / Efficacy | Provide purine nucleoside phosphorylase activity in coupled nucleoside assays. |
| Abbreviation | PNP |
| Customization | Specifications can be customized |
| Specific Activity | >=15.0 U/mg |
| Isoelectric Point | 5.32 |
| Inhibitors | Not inhibited by sodium azide |
| Optimal Temperature | >45 degrees C |
| Thermal Stability | Stable below 70 degrees C |
| Michaelis Constant | 6.4 x 10^-5 M for inosine; 3.2 x 10^-4 M for inorganic phosphate |
| Detection Wavelength | 505 nm |
For research use only, not for clinical use.
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