Proline (Pro) Colorimetric Assay Kit
Proline (Pro) Colorimetric Assay Kit
| SKU # | E-BC-K177-M |
| Detection Instrument | Microplate reader (520 nm) |
| Detection method | Colorimetric method |
Product Details
Properties
| Synonyms | Pro |
| Sample Type | Plant tissue and honey sample |
| Sensitivity | 0.57 μg/mL |
| Detection Range | 0.57-35 μg/mL |
| Detection Method | Colorimetric method |
| Assay type | Quantitative |
| Precision | Average inter-assay CV: 4.7-10.0% | Average intra-assay CV: 3.7-5.5% |
| Other instruments required | Vortex mixer, Water bath, Centrifuge |
| Other reagents required | Acetic acid |
| Storage | 2-8℃ |
| Valid period | 12 months |
Detection Principle
Proline can react with acidic-ninhydrin to form stable red compound. The maximum absorption peak of the compound is at 520 nm. And the concentration of Pro can be calculated by measuring the OD value at 520 nm
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Extracting Solution | 50 mL× 3 vials | 60 mL× 5 vials | 2-8°C, 12 months, shading light |
| Reagent 2 | Ninhydrin | 3 g × 1 vial | 6 g × 1 vial | 2-8°C, 12 months, shading light |
| Reagent 3 | Acid Reagent | 50 mL× 1 vial | 50 mL× 2 vials | 2-8°C, 12 months |
| Reagent 4 | 100 μg/mL Proline Standard |
1 mL× 1 vial | 2 mL× 1 vial | 2-8°C, 12 months, shading light |
| 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 pear 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 (μg/mL) | 3.50 | 12.60 | 30.20 |
| %CV | 5.5 | 5.2 | 3.7 |
Inter-assay Precision
Three human serum samples were assayed 20 times in duplicate by three operators to determine precision between assays.
| Parameters | Sample 1 | Sample 2 | Sample 3 |
| Mean (μg/mL) | 3.50 | 12.60 | 30.20 |
| %CV | 10.0 | 4.7 | 7.5 |
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 100%.
| Standard 1 | Standard 2 | Standard 3 | |
| Expected Conc. (μg/mL) | 8.5 | 18.5 | 28 |
| Observed Conc. (μg/mL) | 8.8 | 17.0 | 29.1 |
| Recovery rate (%) | 104 | 92 | 104 |
Sensitivity
The analytical sensitivity of the assay is 0.57 μg/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.
| Concentration (μg/L) | 0 | 5 | 10 | 15 | 20 | 25 | 30 | 35 |
| Average OD | 0.042 | 0.125 | 0.216 | 0.329 | 0.446 | 0.562 | 0.716 | 0.807 |
| Absoluted OD | 0.000 | 0.084 | 0.174 | 0.288 | 0.404 | 0.520 | 0.650 | 0.765 |