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| Product Name | Catalase Activity Fluorometric Assay Kit |
| Catalog No. | CMTR-HMM-0063 |
| Description | A highly sensitive fluorometric assay for measuring catalase enzyme activity in biological samples. The kit utilizes a proprietary fluorescent probe that reacts with the hydrogen peroxide remaining after catalase-catalyzed decomposition, inversely proportional to catalase activity present in the sample. |
| Intended Use | Quantitative measurement of catalase activity as a key hydrogen peroxide-detoxifying antioxidant enzyme in studies of cellular oxidative stress responses, aging research, metabolic diseases, and evaluation of antioxidant supplement efficacy. |
| Principle / Technology | Catalase rapidly decomposes two molecules of hydrogen peroxide to water and molecular oxygen (2H2O2 → 2H2O + O2) with one of the highest turnover rates of any enzyme (kcat approximately 10^7 s-1); residual H2O2, after exposure to catalase-containing sample for a defined incubation period, is detected by a fluorescent probe that produces a fluorescent product in proportion to H2O2 concentration; catalase activity is quantified by the decrease in fluorescence compared to the no-enzyme control. |
| Detection Method | Fluorescence measurement (Ex/Em: approximately 530/590 nm or compatible green-red channel settings) using a fluorescence microplate reader |
| Sample Type | Cell lysates (mammalian cells, particularly hepatocytes and erythrocytes rich in catalase), tissue homogenates (liver, kidney, red blood cells), purified catalase enzyme, bacterial and fungal extracts |
| Performance Range / Specifications | H2O2 standard curve: 1–50 μM; catalase activity measurable 0.01–10 U/mL range; incubation time 15–30 minutes at room temperature; sample dilution typically 20–500× for tissue samples |
| Sensitivity / LOD | Detection limit: approximately 0.01 U/mL catalase in purified solution; cellular catalase detectable in lysates from approximately 2 × 10^4 cells for high-catalase cell types |
| Specificity | Catalase specifically decomposes hydrogen peroxide; organic peroxides are not substrates; the fluorescent H2O2 detection probe has minimal cross-reactivity with cellular thiols, ascorbate, or transition metal ions under the optimized assay conditions; sodium azide (NaN3) pre-treatment serves as a catalase inhibitor specificity control |
| Reaction Conditions / Protocol | Prepare cell or tissue lysates in cold Assay Buffer (without protease inhibitors that may contain catalase-interfering agents); dilute samples to fall within the linear range of the assay; prepare H2O2 standards and fresh Substrate Solution (H2O2); add 10 μL diluted sample to black microplate wells; add 90 μL Assay Buffer followed by 10 μL H2O2 Substrate Solution; incubate exactly 15 minutes at room temperature; add 50 μL Fluorescent Detection Reagent; incubate additional 15 minutes; measure fluorescence; catalase activity is inversely proportional to fluorescence; calculate activity using the standard curve equation |
| Components / Formulation | Assay Buffer (50 mM potassium phosphate, pH 7.0), H2O2 Substrate Solution (30% H2O2 for dilution), Fluorescent H2O2 Detection Probe (in DMSO), Catalase Positive Control (purified bovine liver catalase, lyophilized), H2O2 Standard (for calibration curve), Sodium Azide inhibitor solution for specificity control |
| Storage Conditions | All kit components at -20°C protected from light except Assay Buffer (2–8°C); Fluorescent Detection Probe sensitive to moisture; reconstituted Catalase standard at -20°C for 4 weeks |
| Shelf Life | 12 months from date of manufacture |
| Package Specifications | 100 assays, 500 assays (96-well format) |
| Product Form | Lyophilized enzyme standard and concentrated liquid detection reagents |
| Quality Control | Each lot tested with catalase standard dilutions for inverse relationship linearity (R2 ≥ 0.98); H2O2 standard curve linearity (R2 ≥ 0.99); sodium azide inhibition >85% of catalase positive control activity |
| Key Features | Fluorometric detection offers 10–50× higher sensitivity than UV absorbance methods; fluorometric probe avoids interference from UV-absorbing biological sample components; includes catalase standard for activity unit calibration; sodium azide inhibitor control validates catalase-specific signal |
For research use only, not for clinical use.
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