Myeloperoxidase (MPO) Activity Assay Kit
SKU: E-BC-K074-S-100
Myeloperoxidase (MPO) Activity Assay Kit
| SKU # | E-BC-K074-S |
| Detection Instrument | Spectrophotometer (460 nm) |
| Detection Method | Colorimetric method |
Product Details
Properties
| Synonyms |
MPO |
| Sample Type | Serum, plasma, milk, animal tissue, cells |
| Sensitivity | 16.95 U/L |
| Detection Range | 16.95-3349 U/L |
| Detection Method | Colorimetric method |
| Assay type | Enzyme Activity |
| Assay time | 90 min |
| Precision | Average inter-assay CV: 9.8% | Average intra-assay CV: 4.5% |
| Other instruments required | Vortex mixer, Micropipettor, Water bath, Incubator, Centrifuge |
| Storage | 2-8℃ |
| Valid period | 12 months |
Dilution of Sample
It is recommended to take 2~3 samples with expected large difference to do pre-experiment before formal experiment and dilute the sample according to the result of the pre-experiment and the detection range (4.9-196.7 U/L).
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| Human serum | 1 |
| Human plasma | 1 |
| Rat serum | 1 |
| 5% Mouse brain tissue homogenization | 1 |
| 5% Mouse heart tissue homogenization | 1 |
|
Human milk |
1 |
Note: The diluent is reagent 2 application solution.
Detection Principle
Myeloperoxidase reduces hydrogen peroxide to a complex. The complex react with o-dianisidine (as hydrogen donor) to produce a yellow product which has a maximum absorption peak at 460 nm. The activity of MPO can be calculated indirectly by measuring the OD value at 460nm.
Kit Components & Storage
| Item | Component | Size 1 (50 assays) | Size 2 (100 assays) | Storage |
| Reagent 1 | Buffer Solution | 18 mL ×1 vial | 36 mL ×1 vial | 2-8℃, 12 months |
| Reagent 2 | Powder A | Powder × 1 vial | Powder × 2 vials | 2-8℃, 12 months |
| Reagent 3 | Powder B | Powder × 1 vial | Powder × 2 vials | 2-8℃, 12 months |
| Reagent 4 | Saline Solution | 10 mL × 1 vial | 10 mL × 2 vials | 2-8℃, 12 months |
| Reagent 5 | Clarificant | 12 mL ×1 vial | 24 mL × 1 vial | 2-8℃, 12 months |
| Reagent 6 | Powder C | Powder × 1 vial | Powder × 2 vials | 2-8℃, 12 months, shading light |
| Reagent 7 | Substrate | 0.15 mL × 1 vial | 0.3 mL × 1 vial | 2-8℃, 12 months |
| Reagent 8 | Acid Reagent | 3 mL × 1 vial | 6 mL × 1 vial | 2-8℃, 12 months |
Note: The reagents must be stored strictly according to the preservation conditions in the above table. The reagents in different kits cannot be mixed with each other. For a small volume of reagents, please centrifuge before use, so as not to obtain sufficient amount of reagents.
Technical Data:
Parameter:
Intra-assay Precision
Three human serum samples were assayed in replicates of 20 to determine precision within an assay (CV = Coefficient of Variation).
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (U/L) | 52.50 | 567.80 | 1126.00 |
| %CV | 4.8 | 4.4 | 4.3 |
Inter-assay Precision
Three human serum samples were assayed 20 times in duplicate by three operators
to determine precision between assays.
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (U/L) | 52.50 | 567.80 | 1126.00 |
| %CV | 10.2 | 9.4 | 9.8 |
Recovery
Take three samples of high concentration, middle concentration and low concentration to test the samples of each concentration for 6 times parallelly to get the average recovery rate of 104%.
| Sample 1 | Sample 2 | Sample 3 | |
| Expected Conc. (U/L) | 684.5 | 1354 | 2686.5 |
| Observed Conc. (U/L) | 711.9 | 1435.2 | 2740.2 |
| Recovery rate (%) | 104 | 106 | 102 |
Sensitivity
The analytical sensitivity of the assay is 16.95 U/L. This was determined by adding two standard deviations to the mean O.D. obtained when the zero standard was assayed 20 times, and calculating the corresponding concentration.