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Peroxidase (POD) Activity Assay Kit (Plant Samples)
SKU: E-BC-K227-S-100
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Peroxidase (POD) Activity Assay Kit (Plant Samples)
| SKU # | E-BC-K227-S |
| Detection Instrument | Spectrophotometer (240 nm, 420 nm) |
| Detection method | Colorimetric method |
Product Details
Properties
| Synonyms | POD |
| Sample Type | Plant tissue |
| Sensitivity | 0.5 U/mL |
| Detection Range | 0.5-40 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 | Incubator, Vortex mixer, Micropipettor, Centrifuge |
| Other reagents required | Normal saline (0.9% NaCl), PBS (0.01 M, pH 7.4) |
| Storage | 2-8℃ |
| Valid period | 12 months |
Images
J Tian et al investigate the cadmium accumulation regulator and the harmful to host plant and leaf bacteria.. Peroxidase (POD) avtivity in crop leaf was determined using peroxidase activity assay kit (E-BC-K227-S).
Activity of POD was significantly induced in OsABCD43-OE9 group. (*P<0.05)
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 (0.5-40 U/mL).
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| 10% Chinese rose tissue homogenization | 1 |
| 10% Epipremnum aureum tissue homogenization | 1 |
| 10% Green pepper tissue homogenization | 2-3 |
| 10% Mushrooms tissue homogenization | 1 |
Note: The diluent is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4).
Detection Principle
The peroxidase can catalyze the decomposition of H2O2 and produce water and oxygen. And oxygen oxidized pyrogallic acid to form yellow product. The activity of peroxidase can be calculated by measuring the absorbance at 420 nm.
Kit Components & Storage
| Item | Component |
Size 1 (50 assays) |
Size 2 (100 assays) |
Storage |
| Reagent 1 | Buffer Solution | 60 mL × 2 vials | 60 mL × 4 vials | 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 | 1.5 mL × 2 vials | 1.5 mL × 4 vials | 2-8℃, 12 months |
| Reagent 4 | Stop Solution | 30 mL × 1 vial | 60 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/mL) | 6.20 | 18.40 | 30.50 |
| %CV | 4.1 | 3.7 | 3.6 |
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) | 6.20 | 18.40 | 30.50 |
| %CV | 5.2 | 5.6 | 5.7 |
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 98%.
| Sample 1 | Sample 2 | Sample 3 | |
| Expected Conc. (U/mL) | 12.4 | 21.5 | 35 |
| Observed Conc. (U/mL) | 12.3 | 21.7 | 32.9 |
| Recovery rate (%) | 99 | 101 | 94 |
Sensitivity
The analytical sensitivity of the assay is 0.5 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:
| Activity (U/mL) | 1.375 | 0.69 | 0.344 | 0.172 | 0.086 | 0.043 | 0.022 | 0.011 | 0.005 | 0 |
| Average OD | 0.655 | 0.649 | 0.624 | 0.461 | 0.293 | 0.250 | 0.109 | 0.074 | 0.051 | 0.043 |
| Absoluted OD | 0.612 | 0.649 | 0.581 | 0.461 | 0.250 | 0.130 | 0.109 | 0.031 | 0.008 | 0 |