Phosphoenolpyruvate Carboxylase (PEPC) Activity Assay Kit
Phosphoenolpyruvate Carboxylase (PEPC) Activity Assay Kit
| SKU # | E-BC-K613-M |
| Detection Instrument | Microplate reader (320-360 nm, optimum wavelength: 340 nm) |
| Detection method | Colorimetric method |
Product Details
Properties
| Synonyms | PEPC |
| Sample type | Plant tissue |
| Sensitivity | 0.72 U/L |
| Detection range | 0.72-25.61 U/L |
| Detection Method | Colorimetric method |
| Assay type | Enzyme Activity |
| Precision | Average inter-assay CV: 8.5-10.6% | Average intra-assay CV: 1.5-4.3% |
| Storage | 2-8℃ |
| Valid period | 12 months |
Dilution of Sample
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| 10% Shanghai green tissue homogenate | 1 |
| 10% Sweet potato leaves tissue homogenate | 1 |
| 10% Crowndaisy chrysanthemum tissue homogenate | 1 |
| 10% Napa cabbage tissue homogenate | 1 |
Note: The diluent is PBS (0.01 M, pH 7.4). For the dilution of other sample types, please do pretest to confirm the dilution factor.
Detection Principle
Phosphoenolpyruvate carboxylase (PEPC) catalyzes phosphoenolpyruvate (PEP) to produce oxaloacetic acid, which further reacts with NADH. NADH has maximum absorption at 340 nm. The activity of PEPC can be calculated by measuring the change of absorbance value at 340 nm.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Buffer Solution | 13 mL ×1 vial | 25 mL × 1 vial | 2-8℃, 12 months, shading light |
| Reagent 2 | Substrate | Powder × 2 vials | Powder × 4 vials | 2-8℃, 12 months, shading light |
| UV 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 shanghai green tissue 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) | 3.60 | 11.80 | 19.50 |
| %CV | 4.3 | 2.9 | 1.5 |
Inter-assay Precision
Three shanghai green tissue 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) | 3.60 | 11.80 | 19.50 |
| %CV | 10.6 | 8.5 | 9.4 |
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%.
| Sample 1 | Sample 2 | Sample 3 | |
| Expected Conc. (U/L) | 7.6 | 15.5 | 20 |
| Observed Conc. (U/L) | 8.4 | 14.9 | 20.4 |
| Recovery rate (%) | 111 | 9.6 | 102 |
Sensitivity
The analytical sensitivity of the assay is 0.72 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.