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| Product Name | >=25 U/mg Pyruvate Kinase for 340 nm Potassium Assay Systems |
| Catalog No. | IEC-HMM-0247 |
| Description | Alta DiagnoTech pyruvate kinase is a pale-yellowish white amorphous lyophilized enzyme powder for potassium biochemical reagent development. |
| Intended Use | Enzyme raw material for potassium biochemical assay reagents. |
| Detection Method | LDH-coupled kinetic absorbance assay at 340 nm |
| Performance Range / Specifications | Specific activity >=25.0 U/mg; assay linear range 300-1,000 U/L. |
| Reaction Conditions / Protocol | Prepare Reagent 1 by successively dissolving 18.2 g triethanolamine, 5.4 g magnesium chloride hexahydrate, 3.24 g potassium chloride, 8.8 g ADP potassium salt and 8.06 g phosphoenolpyruvate tricyclohexylammonium salt in 800 mL water; adjust to pH 7.6 at 25 degrees C with HCl, dissolve 10 kU lactate dehydrogenase and bring to 1 L. Complete preparation within 1 hour and testing within 4 hours. Prepare Reagent 2 by dissolving 4.84 g Tris in 800 mL water, adjusting to pH 9.0 at 25 degrees C with HCl, adding 0.8 g NADH and bringing to 1 L; sterile-filter through 0.22 um and store for up to 2 weeks at 2-8 degrees C. Combine 0.900 mL Reagent 1 and 0.300 mL Reagent 2, preincubate for 90 seconds at 37 degrees C, add 0.040 mL sample, pre-equilibrate for 160 seconds and measure the 1-minute absorbance-change rate at 340 nm. Activity in U/L equals Delta A/min x 4,984 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 lyophilized powder |
| Activity / Unit Definition | One unit is the amount of enzyme that oxidizes 1 umol NADH per minute under the stated reaction conditions. |
| Molecular Weight | Approximately 62 kDa |
| pH Range / Optimal pH | Stable at pH 5.0-9.5; optimum pH 8.0 |
| Recommended Buffer System | Sample diluent: 50 mM Tris-HCl at pH 7.5 containing 1 g/L BSA. Dissolve the weighed enzyme sample in an ice-water bath. |
| Appearance | Pale-yellowish white amorphous lyophilized powder |
| Alias | Pyruvate Kinase |
| Product Type | Pyruvate kinase enzyme raw material |
| Function / Efficacy | Catalyze phosphoenolpyruvate-dependent ATP formation in coupled potassium assays. |
| Abbreviation | PK |
| Customization | Specifications can be customized |
| Specific Activity | >=25.0 U/mg |
| Isoelectric Point | 5.71 |
| Inhibitors | Not inhibited by sodium azide |
| Optimal Temperature | 50 degrees C |
| Thermal Stability | Stable below 50 degrees C |
| Michaelis Constant | 3.6 x 10^-4 M for ADP potassium salt; 9.1 x 10^-4 M for phosphoenolpyruvate potassium salt |
| Activator | Mg2+ and K+ |
| Detection Wavelength | 340 nm |
| Linear Range | 300-1,000 U/L |
For research use only, not for clinical use.
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