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Catalase (CAT) Activity Assay Kit
SKU: E-BC-K031-M-96
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Catalase (CAT) Activity Assay Kit
| SKU # | E-BC-K031-M |
| Detection Instrument | Microplate reader (400-410 nm, optimum wavelength: 405 nm) |
| Detection Method | Colorimetric method |
Product Details
Properties
| Synonyms | CAT |
| Sample Type | Serum, plasma, cells, cell culture supernatant, tissue |
| Sensitivity | 1.12 U/mL |
| Detection Range | 1.12 -150 U/mL |
| Detection Method | Colorimetric method |
| Assay type | Enzyme Activity |
| Assay time | 40 min |
| Precision | Average inter-assay CV: 7.700% | Average intra-assay CV: 3.900% |
| Other instruments required | Micropipettor, Vortex Mixer, Incubator |
| 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 (1.12 -150 U/mL).
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
| Human serum | 1 |
| 293T supernatant | 1 |
| 10% Rat heart tissue homogenization | 50-100 |
| 10% Rat liver tissue homogenization | 100-200 |
| 10% Rat spleen tissue homogenization | 50-100 |
| Mouse serum | 1 |
| 10% Epipremnum aureum tissue homogenization | 1-2 |
| 10% Rat lung tissue homogenization | 50-100 |
| 10% Rat kidney tissue homogenization | 50-100 |
| 10% Rat brain tissue homogenization | 20-50 |
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(48 T) | Size 2(96 T) | Storage |
| Reagent 1 | Buffer Solution | 12 mL × 1 vial | 24 mL × 1 vial | 2-8℃, 12 months |
| Reagent 2 | Substrate | 1.5 mL × 1 vial | 1.5 mL × 2 vials | 2-8℃, 12 months |
| Reagent 3 | Chromogenic Agent | Powder ×1 vial | Powder ×1 vial | 2-8℃, 12 months |
| Reagent 4 | Clarificant | 1.5 mL × 1 vial | 1.5 mL × 2 vials | 2-8℃, 12 months |
| Reagent 5 | 9.6 mol/L H2O2 Standard Solution | 1.5 mL× 1 vial | 1.5 mL × 2 vials | 2-8℃, 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 (U/mL) | 3.70 | 26.40 | 78.60 |
| %CV | 4.2 | 3.8 | 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 (U/mL) | 3.70 | 26.40 | 78.60 |
| %CV | 7.4 | 7.9 | 7.8 |
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. (μmol/mL) | 15 | 36 | 54 |
| Observed Conc. (μmol/mL) | 15.2 | 34.9 | 55.1 |
| Recovery rate (%) | 101 | 97 | 102 |
Sensitivity
The analytical sensitivity of the assay is 1.12 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.
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 (μmol/mL) | 0 | 10 | 20 | 40 | 50 | 60 | 80 | 100 |
| Average OD | 0.062 | 0.087 | 0.118 | 0.143 | 0.169 | 0.192 | 0.219 | 0.311 |
| Absoluted OD | 0 | 0.025 | 0.056 | 0.081 | 0.107 | 0.13 | 0.157 | 0.249 |