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| Product Name | >=1.5 U/mg Grade III Pyruvate Oxidase for Pyruvate, GOT and GPT Determination |
| Catalog No. | IEC-HMM-0248 |
| Description | Alta DiagnoTech pyruvate oxidase is a yellowish lyophilized amorphous powder used for enzymatic determination of pyruvate and transaminase activity. |
| Intended Use | Enzyme raw material for enzymatic determination of pyruvate, GOT and GPT in clinical-analysis reagent development. |
| Principle / Technology | With FAD, TPP and Mg2+, pyruvate oxidase converts pyruvate, phosphate and oxygen to acetyl phosphate, carbon dioxide and hydrogen peroxide. A coupled peroxidase reaction forms a quinoneimine dye for photometric measurement. |
| Detection Method | Coupled colorimetric enzyme assay at 550 nm |
| Specificity | Relative activity at 50 mM substrate: pyruvate 100%, alpha-ketobutyrate 5.8%, and 0% for alpha-ketoglutarate, oxaloacetate, DL-lactate, acetate, acetoacetate, L-alanine and L-aspartate. |
| Reaction Conditions / Protocol | The assay mixture contains 48 mM pyruvate, 50 mM potassium phosphate, 0.48 mM 4-aminoantipyrine, 0.58 mM EHSP, 0.019 mM TPP, 0.01 mM FAD, 0.97 mM EDTA, 9.7 mM MgSO4 and approximately 4.8 U/mL peroxidase at pH 5.9. Equilibrate 2.5 mL mixture plus 0.5 mL pyruvate solution for about 5 minutes at 37 degrees C, add 0.1 mL enzyme, then record A550 for 3-4 minutes. |
| Storage Conditions | Store at -20 degrees C |
| Shelf Life | At least 1 year at -20 degrees C |
| Product Form | Yellowish lyophilized amorphous powder |
| Purity | Contaminating ATPase <=5.0 x 10^-2%; GOT and GPT each <=5.0 x 10^-2% |
| Activity / Unit Definition | One unit forms 1 umol hydrogen peroxide, equivalent to 0.5 umol quinoneimine dye, per minute under the stated assay conditions. |
| Molecular Weight | Approximately 260 kDa |
| pH Range / Optimal pH | Stable at pH 5.7-6.5 for 20 hours at 25 degrees C; optimum pH 5.7 |
| Appearance | Yellowish amorphous lyophilized powder |
| Alias | Pyruvate Oxidase |
| Product Type | Pyruvate oxidase enzyme raw material |
| Function / Efficacy | Oxidize pyruvate while generating hydrogen peroxide for coupled photometric assays. |
| Abbreviation | POX |
| Customization | Specifications can be customized |
| EC Number | EC 1.2.3.3 |
| Specific Activity | >=1.5 U/mg solid, Grade III |
| Isoelectric Point | 4.3 |
| Inhibitors | Fe3+, Zn2+, Cu2+, Ag+ and Hg2+ |
| Optimal Temperature | 65 degrees C |
| Thermal Stability | Stable below 45 degrees C for 15 minutes at pH 6.0 |
| Michaelis Constant | 3.4 x 10^-4 M for pyruvate |
| Stabilizer | Sugars and FAD |
| Detection Wavelength | 550 nm |
| Interference | Residual activity after 1 hour at 25 degrees C in 50 mM potassium phosphate, pH 6.0: none 100%; 20 mM MgCl2 95%, CaCl2 93%, Ba(OAc)2 97%, CoCl2 84%, MnCl2 76%, ZnSO4 48%, Cd(OAc)2 86%, NiCl2 119%, CuSO4 0.9%, Pb(OAc)2 33%, NaF 100%, NaN3 94%; 2 mM FeCl3 8.4%, HgCl2 0%, o-phenanthroline 97%, IAA 102%, NEM 104%, MIA 96%; 5 mM EDTA 107%; 10 mM alpha,alpha'-dipyridyl 95%; 50 mM borate 102%; 1 mM PCMB 66%; 20 mM hydroxylamine 98%; 0.10% Triton X-100 143%, Brij 35 133%, Tween 20 146%, Span 20 121%, sodium cholate 116%; 0.05% SDS 85% and DAC 53%. |
For research use only, not for clinical use.
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