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Myeloperoxidase (MPO) Peroxidation Activity Fluorometric Assay Kit
SKU: E-BC-F013-500
Myeloperoxidase (MPO) Peroxidation Activity Fluorometric Assay Kit
| SKU # | E-BC-F013 |
| Detection Instrument | Fluorescence microplate reader (Ex/Em=535 nm/587 nm) |
| Detection Method | Fluorescence method |
Product Details
Properties
| Synonyms | MPO |
| Sample Type | Serum, plasma, animal tissue |
| Sensitivity | 0.001 U/L |
| Detection Range | 0.001 - 1.26 U/L |
| Detection Method | Fluorescence method |
| Assay Time | 20 min |
| Precision | Inter-assay CV: 5.4% | intra-assay CV: 1% |
| Storage | This product can be stored at -20°C for 12 months with shading light. |
| Valid period | 12 months |
Dilution Of Sample
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
|---|---|
| Porcine serum | 5-10 |
| Rabbit serum | 3-5 |
| Rat serum | 2-5 |
| Mouse serum | 10-20 |
| Mouse plasma | 30-50 |
| Horse serum | 2-5 |
| 10% Rat heart tissue homogenate | 1 |
| 10% Rat lung tissue homogenate | 1 |
The diluent is buffer solution. For the dilution of other sample types, please do pretest to confirm the dilution factor.
Detection Principle
Under the catalysis of peroxidase, hydrogen peroxide can oxidize the non-fluorescent probe into the fluorescent substance, and its fluorescence intensity is proportional to the total peroxidase activity in the sample. This kit specifically inhibits the peroxidase activity of MPO in the sample through an MPO enzyme inhibitor, thus distinguishing the peroxidase activity of MPO in the sample from that of other peroxidases.
Kit Components & Storage
| Item | Component | Size (96 T) | Storage |
| Reagent 1 | Buffer Solution | 60 mL × 1 vial | 2-8°C, 12 months |
| Reagent 2 | Probe | 0.25 mL × 1 vial | 2-8°C, 12 months shading light |
| Reagent 3 | Substrate | 0.25 mL × 1 vial | 2-8°C, 12 months |
| Reagent 4 | Inhibitor | 1.2 mL × 1 vial | 2-8°C, 12 months |
| Reagent 5 | 25 μmol/L Resorufin Standard | 1.5 mL × 1 vial | -20℃, 12 months, shading light |
| Black Microplate | 96 wells | No requirement | |
| Plate Sealer | 2 pieces |
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) | 0.01 | 0.30 | 1.00 |
| %CV | 1.2 | 0.9 | 0.9 |
Inter-assay Precision
Three human serum samples were assayed 20times in duplicate by three operators to determine precision between assays.
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (U/L) | 0.01 | 0.30 | 1.00 |
| %CV | 5.2 | 4.8 | 6.2 |
Sensitivity
The analytical sensitivity of the assay is 0.001 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.
Standard Curve:
As the fluorescence value of the standard curve may vary according to the conditions of the actual assay performance (e.g. operator, pipetting technique or temperature effects), so the standard curve and data are provided as below for reference only:
| Concentration (μmol/L) | 0 | 2 | 4 | 6 | 8 | 10 | 12 | 15 |
| Fluorescence value | 28 | 1055 | 2096 | 3079 | 4118 | 4973 | 4973 | 6717 |
| 28 | 1079 | 2110 | 2904 | 3893 | 5233 | 5923 | 7063 | |
| Average fluorescence value | 28 | 1067 | 2103 | 2992 | 4006 | 5103 | 5802 | 6890 |
| Absoluted fluorescence value | 0 | 1039 | 2075 | 2992 | 3978 | 5075 | 5774 | 6862 |