Protein Carbonyl Colorimetric Assay Kit (Tissue and Serum Samples)
SKU: E-BC-K117-S-100
Protein Carbonyl Colorimetric Assay Kit (Tissue and Serum Samples)
| SKU # | E-BC-K117-S |
| Detection Instrument | Spectrophotometer (370 nm) |
| Detection Method | Colorimetric method |
Product Details
Properties
| Sample Type | Serum, plasma, hydrothorax, cell culture supernatant, tissue |
| Sensitivity | 0.02 nmol/mgprot |
| Detection range | 0.02-10 nmol/mgprot |
| Detection Method | Colorimetric method |
| Assay type | Quantitative |
| Assay time | 240 min |
| Precision | Average inter-assay CV: 8.600% | Average intra-assay CV: 4.500% |
| Other instruments required | Vortex mixer, Micropipettor, Water bath, Incubator, Centrifuge |
| Other reagents required |
Absolute ethanol, Ethyl acetate |
| Storage | 2-8℃ |
| Valid period | 12 months |
Dilution of Sample
It is recommended to take 2~3 samples with expected large difference to do pre-experiment before formal experiment. The sample can be detected by this kit when the protein content of samples is ranged from 1-10 mg/mL.
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| Human serum | 8-10 |
| Mouse serum | 8-10 |
| 10% Rat liver tissue homogenization | 2-3 |
| 10% Mouse heart tissue homogenization | 1 |
| 10% Mouse brain tissue homogenization | 1 |
Note: The diluent is double distilled water or reagent 1.
Detection Principle
The content of protein carbonyl increased after oxidation, and the carbonyl group reacted with 2, 4-dinitrophenylhydrazine to form a reddish brown precipitate. The absorbance can be measured at 370 nm after the precipitation is dissolved. The carbonyl content can be calculated indirectly.
Kit Components & Storage
| Item | Component |
Size 1 (50 Assays) |
Size 2 (100 Assays) |
Storage |
| Reagent 1 | Homogenate Medium | 50 mL ×1 vial | 50 mL × 2 vials | 2-8°C, 12 months |
| Reagent 2 | Sulfates | Powder ×1 vial | Powder ×2 vials | 2-8°C, 12 months, shading light |
| Reagent 3 | DNPH Solution | 10 mL ×1 vial | 20 mL ×1 vial | 2-8°C, 12 months, shading light |
| Reagent 4 | Acid Reagent | 10 mL ×1 vial | 20 mL ×1 vial | 2-8°C, 12 months |
| Reagent 5 | Protein Precipitator | 30 mL ×1 vial | 60 mL ×1 vial | 2-8°C, 12 months |
| Reagent 6 | Denaturant | 37.5 mL ×2 vials | 50 mL ×3 vials | 2-8°C, 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 (nmol/mgprot) | 1.50 | 3.80 | 7.90 |
| %CV | 4.8 | 4.3 | 4.4 |
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 (nmol/mgprot) | 1.50 | 3.80 | 7.90 |
| %CV | 8.4 | 8.5 | 8.9 |
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. (nmol/mgprot) | 2.4 | 5.2 | 8.5 |
| Observed Conc. (nmol/mgprot) | 2.4 | 5.4 | 8.6 |
| Recovery rate (%) | 99 | 103 | 101 |
Sensitivity
The analytical sensitivity of the assay is 0.02 nmol/mgprot. 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.