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Peroxidase (POD) Activity Assay Kit (Plant Samples)
SKU: E-BC-K227-M-96
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Peroxidase (POD) Activity Assay Kit (Plant Samples)
| SKU # | E-BC-K227-M |
| Detection Instrument | Microplate reader (410 nm-430 nm, optimum wavelength: 420 nm) |
| Detection method | Colorimetric method |
Product Details
Properties
| Synonyms | POD |
| Sample Type | Plant tissue |
| Sensitivity | 0.01 U/mL |
| Detection Range | 0.01–100 U/mL |
| Detection Method | Colorimetric method |
| Assay type | Enzyme Activity |
| Assay time | 60 min |
| Precision | Average inter-assay CV: 5% | Average intra-assay CV: 3.200% |
| Other instruments required | Spectrophotometer (240 nm), Micropipettor, Vortex mixer, Centrifuge, 37℃ Incubator |
| Other reagents required | PBS (0.01 M, pH 7.4) |
| 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 (0.01-100 U/mL).
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| 10% Green pepper tissue homogenate | 1 |
| 10% Chive leaf tissue homogenate | 1 |
| 10% Photinia leaf tissue homogenate | 1 |
| 10% Epipremnum aureum tissue homogenate | 1 |
| 10% Mushrooms tissue homogenate | 1 |
| 10% White radish tissue homogenate | 1 |
Note: The diluent is PBS (0.01 M, pH 7.4).
Detection Principle
Plant peroxidase, a member of the superfamily of peroxidase, catalyzes the redox reaction between H2O2 and various reductants. The plant peroxidase has the same general structure and consists of iron porphyrin IX and ten α-helixes. Based on the difference of primary structure, the superfamily of plant peroxidase can be divided into three types: class I (intracellular type), class II (extracellular type of fungi) and class III (secreted type of plant).
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Buffer Solution | 30 mL × 1 vial | 60 mL × 1 vial | 2-8℃, 12 months |
| Reagent 2 | Chromogenic Agent | Powder × 1 vial | Powder × 2 vials | 2-8°C, 12 months, shading light |
| Reagent 3 | Substrate Solution | 1 mL × 1 vial | 1 mL × 1 vial | 2-8℃, 12 months |
| Reagent 4 | Stop Solution | 10 mL × 1 vial | 20 mL × 1 vial | 2-8℃, 12 months |
| 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/mL) | 5.80 | 36.20 | 75.00 |
| %CV | 3.6 | 3.1 | 2.9 |
Inter-assay Precision
Three human serum samples were assayed 17 times in duplicate by three operators to determine precision between assays.
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (U/mL) | 5.80 | 36.20 | 75.00 |
| %CV | 3.6 | 3.1 | 8.3 |
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 102%.
| Sample 1 | Sample 2 | Sample 3 | |
| Expected Conc. (U/mL) | 12.5 | 47.6 | 88.5 |
| Observed Conc. (U/mL) | 12.4 | 50.0 | 90.3 |
| Recovery rate (%) | 99 | 105 | 102 |
Sensitivity
The analytical sensitivity of the assay is 0.01 U/mL. 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 OD 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: