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Catalase (CAT) Activity Assay Kit
SKU: E-BC-K031-S-100
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Catalase (CAT) Activity Assay Kit
| SKU # | E-BC-K031-S |
| Detection Instrument | Spectrophotometer(405 nm) |
| Detection Method | Colorimetric method |
Product Details
Properties
| Synonyms | CAT |
| Sample Type | Serum, plasma, cells, tissue |
| Sensitivity | 0.27 U/mL |
| Detection Range | 0.27-155.4 U/mL |
| Detection Method | Colorimetric method |
| Assay type | Enzyme Activity |
| Assay time | 70 min |
| Precision | Average inter-assay CV: 5.100% | Average intra-assay CV: 3.100% |
| Other instruments required | Micropipettor, Vortex mixer, Incubator or Water-bath |
| Other reagents required | Normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4) |
| 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 and dilute the sample according to the result of the pre-experiment and the detection range (0.27-155.4 U/mL).
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor | The volume of sample |
| 10% Rat liver tissue homogenization | 25-50 | 50 μL |
| 10% Rat kidney tissue homogenization | 10-25 | 50 μL |
| 10% Rat brain tissue homogenization | 05-10 | 50 μL |
| Human serum | 1 | 100 μL |
| 293T supernatant | 1 | 100 μL |
Note: The diluent is normal saline (0.9% NaCl) or PBS (0.01 M, pH 7.4).
Detection Principle
The reaction that catalase (CAT) decomposes H2O2 can be quickly stopped by ammonium molybdate. The residual H2O2 reacts with ammonium molybdate to generate a yellowish complex. CAT activity can be calculated by production of the yellowish complex at 405 nm.
Kit Components & Storage
| Item | Component | Size 1 (50 assays) | Size 2 (100 assays) | Storage |
| Reagent 1 | Buffer Solution | 60 mL × 1 vial | 60 mL × 2 vials | 2-8℃, 12 months |
| Reagent 2 | Substrate | 12 mL × 1 vial | 12 mL × 1 vial | 2-8℃, 12 months |
| Reagent 3 | Chromogenic Agent | Powder ×1 vial | Powder × 2 vials | 2-8℃, 12 months |
| Reagent 4 | Clarificant | 6 mL × 1 vial | 12 mL × 1 vial | 2-8℃, 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 (U/mL) | 3.50 | 34.50 | 88.50 |
| %CV | 3.6 | 3.2 | 2.5 |
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 (U/mL) | 3.50 | 34.50 | 88.50 |
| %CV | 5.7 | 4.9 | 4.7 |
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 96%
| Sample 1 | Sample 2 | Sample 3 | |
| Expected Conc. (U/mL) | 19 | 77 | 106 |
| Observed Conc. (U/mL) | 18.4 | 73.2 | 101.8 |
| Recovery rate (%) | 97 | 95 | 96 |
Sensitivity
The analytical sensitivity of the assay is 0.27 U/mL CAT. 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.