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Urine Protein Colorimetric Assay Kit
SKU: E-BC-K252-M-96
Urine Protein Colorimetric Assay Kit
| SKU # | E-BC-K252-M |
| Detection Instrument | Microplate reader (585-605 nm, optimum wavelength: 595 nm) |
| Detection method | Colorimetric method |
Product Details
Properties
| Sample type | Urine |
| Sensitivity | 7.77 mg/L |
| Detection Range | 7.77-100 mg/L |
| Detection Method | Colorimetric method |
| Assay type | Quantitative |
| Assay time | 15 min |
| Precision | Average inter-assay CV: 4.700% | Average intra-assay CV: 2.400% |
| Other instruments required | Centrifuge, |
| Storage | -20℃ |
| Valid period | 12 months |
Detection Principle
The protein in urine reacts with Coomassie Blue G-250 under acidic conditions to produce chromogenic substance. The reaction system changes from brownish red to blue. The blue substance produced has the maximum absorption wavelength at 585-605 nm.
Kit Components & Storage
| Item | Component | Size 1(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Chromogenic Agent | 12 mL × 1 vial | 24 mL × 1 vial | -20°C, 12 months shading light |
| Reagent 2 | 500 mg/L Standard Solution | 0.5 mL × 1 vial | 1 mL × 1 vial | -20°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 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 (mg/L) | 15.00 | 42.00 | 85.00 |
| %CV | 2.6 | 2.4 | 2.2 |
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 (mg/L) | 15.00 | 42.00 | 85.00 |
| %CV | 4.3 | 4.7 | 5.1 |
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 105%.
| Parameters | Standard 1 | Standard 2 | Standard 3 |
| Expected Conc. (mg/L) | 35 | 65 | 88 |
| Observed Conc. (mg/L) | 35.4 | 69.6 | 94.2 |
| Recovery rate (%) | 101 | 107 | 107 |
Sensitivity
The analytical sensitivity of the assay is 7.77 mg/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.
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 (mg/L) |
0 | 30 | 40 | 60 | 70 | 80 | 90 | 100 |
| OD value | 0.290 | 0.340 | 0.359 | 0.385 | 0.412 | 0.420 | 0.447 | 0.480 |
| 0.296 | 0.343 | 0.363 | 0.405 | 0.425 | 0.440 | 0.460 | 0.492 | |
| Average OD | 0.293 | 0.342 | 0.361 | 0.395 | 0.419 | 0.430 | 0.454 | 0.486 |
| Absoluted OD | 0.000 | 0.049 | 0.068 | 0.102 | 0.126 | 0.137 | 0.161 | 0.193 |