Plant Root Activity Assay Kit
Plant Root Activity Assay Kit
| SKU # | E-BC-K855-S |
| Detection Instrument | Spectrophotometer (485 nm) |
|
Detection method |
Colorimetric method |
Product Details
Properties
| Synonyms | Plant tissue |
| Sensitivity | 0 U/g wet weight |
| Detection range | 0-0.67 U/g wet weight |
| Detection method | Colorimetric method |
| Assay type | Enzyme Activity |
| Assay time | 3.5h |
| Precision | Average inter-assay CV: 3% | Average intra-assay CV: 1% |
| Other instruments required | Centrifuge, Incubator |
| Other reagents required | Double distilled water, Ethyl Acetate |
| Storage | -20℃ |
| Valid period | 12 months |
Dilution of Sample
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| Cymbidium | 1 |
| Black nightshade | 1 |
| Stone step grass | 1 |
| Bitter fleabane | 1 |
| Hemlock chervil | 1 |
| Oilseed rape | 1 |
Note: The diluent is ethyl acetate. For the dilution of other sample types, please do pretest to confirm the dilution factor
Detection Principle
The detection principle of the kit is to use 2，3，5-triphenyltetrazolium chloride (TTC) as the substrate, incubate for 1 to 3 hours, the dehydrogenase in the root can reduce TTC and generate 1,3,5-tiphenylformazan (TTF) which is insoluble in water, then TTF will be extracted from the root with organic solvent (ethyl acetate or acetone, etc.) By detecting the absorbance at 485nm, the amount of TTC reduction can be calculated, which represents the dehydrogenase activity and is used as an index of plant root activity. The kit is used for quantitative determination of plant root activity or dehydrogenase activity.
Kit Components & Storage
| Item | Component | Size (100 Assays) | Storage |
| Reagent 1 | Buffer Solution | Power × 1 vial | -20℃, 12 months |
| Reagent 2 | Chromogenic Agent | Power × 1 vial | -20℃, 12 months, shading light |
| Reagent 3 | Reducing Reagent | 12 mL×1 vial | -20℃, 12 months, shading light |
| Reagent 4 | Stop Solution | Power × 1 vial |
-20℃, 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 cymbidium 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/g wet weight) | 0.08 | 0.24 | 0.52 |
| %CV | 1.2 | 0.9 | 0.9 |
Inter-assay Precision
Three cymbidium samples were assayed 20 times in duplicate by three operators to determine precision between assays.
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (U/g wet weight) | 0.08 | 0.24 | 0.52 |
| %CV | 2.7 | 3.2 | 3.1 |
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 103%.
| Standard 1 | Standard 2 | Standard 3 | |
| Expected Conc. (μg) | 35 | 95 | 163 |
| Observed Conc. (μg) | 35.4 | 98.8 | 169.5 |
| Recovery rate(%) | 101 | 104 | 104 |
Sensitivity
The analytical sensitivity of the assay is 0 U/g wet weight. 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:
| Standard quality (μg) | 0 | 25 | 50 | 75 | 100 | 150 | 200 |
| OD Value | 0 | 0.134 | 0.248 | 0.340 | 0.380 | 0.644 | 0.872 |