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Catalase (CAT) Activity Fluorometric Assay Kit
SKU: E-BC-F006-500
Catalase (CAT) Activity Fluorometric Assay Kit
| SKU # | E-BC-F006 |
| Detection Instrument | Fluorescence microplate reader (Ex/Em=535 nm/587 nm) |
| Detection Method | Fluorescence method |
Product Details
Properties
| Synonyms | CAT |
| Sample type | Serum, plasma, animal tissue |
| Sensitivity | 0.01 U/L |
| Detection Range | 0.01-6.51 U/L |
| Detection Method | Fluorescence method |
| Assay Time | 30 min |
| Precision | Inter-assay CV: 6.8% | intra-assay CV: 3.4% |
| Storage | This product can be stored at -20°C for 12 months with shading light. |
| Valid period | 12 months |
Dilution Of Sample
The recommended dilution factor for different samples is as follows (for reference only):
| Sample type | Dilution factor |
|---|---|
| Mouse serum | 60-80 |
| Mouse plasma | 60-80 |
| Rat serum | 70-80 |
| Human saliva | 30-50 |
| Rat urine | 30-50 |
| 10% Mouse liver tissue homogenate | 2500-3000 |
| 10% Mouse lung tissue homogenate | 200-400 |
| 10% Rat muscle tissue homogenate | 100-200 |
| 10% Rat brain tissue homogenate | 40-50 |
| 10% Mouse kidney tissue homogenate | 2000-2500 |
Detection Principle
Catalase can decompose H2O2 to generate H2O and O2, the residual H2O2 in the detection system react with the fluorescent substance, and the content of residual H2O2 is proportional to the fluorescence intensity at the excitation wavelength of 535 nm and emission wavelength of 587 nm, the catalase activity is inversely proportional to the fluorescence intensity.
Kit Components & Storage
| Item | Component | Size (96 T) | Storage |
| Reagent 1 | Buffer Solution | 60 mL ×1 vial | 20°C, 12 months |
| Reagent 2 | Substrate | 0.1 mL × 1 vial | -20°C, 12 months shading light |
| Reagent 3 | Probe Solution | 0.12 mL × 1 vial | -20°C, 12 months shading light |
| Reagent 4 | Enzyme Reagent | Powder × 1 vial | -20°C, 12 months shading light |
| Reagent 5 | 1 mol/L H2O2 Standard Solution | 0.4 mL × 1 vial | -20°C, 12 months shading light |
| Black 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/L) | 0.80 | 3.00 | 5.40 |
| %CV | 4.2 | 3.5 | 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/L) | 0.80 | 3.00 | 5.40 |
| %CV | 7.5 | 6.0 | 6.9 |
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 92%.
| Standard 1 | Standard 2 | Standard 3 | |
| Expected Conc.(μmol/L) | 3 | 25 | 38 |
| Observed Conc.(μmol/L) | 2.7 | 23.3 | 35.3 |
| Recovery Rate (%) | 90 | 93 | 93 |
Sensitivity
The analytical sensitivity of the assay is 0.01 U/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 (μmol/L) | 0 | 1 | 5 | 10 | 20 | 30 | 40 | 50 |
| Fluorescence value | 102 | 351 | 1209 | 2542 | 5341 | 7984 | 10023 | 13847 |
| 101 | 351 | 1229 | 2603 | 5434 | 8133 | 10396 | 13778 | |
| Average fluorescence value | 102 | 351 | 1219 | 2572 | 5387 | 8058 | 10209 | 13812 |
| Absoluted fluorescence value | 0 | 249 | 1117 | 2471 | 5286 | 7957 | 10108 | 13711 |