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| Product Name | Superoxide Dismutase (SOD) Activity Colorimetric Assay Kit |
| Catalog No. | CMTR-HMM-0062 |
| Description | A colorimetric assay kit for measuring superoxide dismutase enzyme activity in biological samples based on the inhibition of WST-1 reduction by superoxide anions generated by the xanthine/xanthine oxidase system. SOD present in the sample competes for superoxide, reducing WST-1 formazan formation in proportion to enzyme activity. |
| Intended Use | Quantification of total SOD activity (Cu/Zn-SOD, Mn-SOD, and Fe-SOD) in cell lysates, tissue homogenates, and biological fluids as an indicator of cellular antioxidant defense capacity in oxidative stress research. |
| Principle / Technology | Xanthine oxidase catalyzes the oxidation of xanthine to uric acid with concomitant production of superoxide radical (O2•-); superoxide reduces the water-soluble tetrazolium salt WST-1 to a yellow formazan product (absorbance 450 nm); SOD in the sample catalyzes the dismutation of superoxide to H2O2 and O2, competing with WST-1 and thereby reducing formazan generation; one unit of SOD activity is defined as the amount of enzyme causing 50% inhibition of the WST-1 reduction rate. |
| Detection Method | Colorimetric kinetic absorbance measurement at 450 nm over 20–30 minutes using a microplate spectrophotometer; inhibition rate of WST-1 reduction is proportional to SOD activity |
| Sample Type | Cell lysates (cultured mammalian cells), tissue homogenates (liver, kidney, heart, brain), erythrocyte lysates, serum and plasma, plant tissue extracts, bacterial lysates |
| Performance Range / Specifications | SOD standard activity range: 0.1–20 U/mL; sample dilution typically 10–100× depending on SOD content; kinetic reading over 20 minutes at 37°C; 50% inhibition of WST-1 reduction rate defines the linear quantitation range |
| Sensitivity / LOD | Detection limit: approximately 0.1 U/mL SOD activity in purified enzyme preparation; cellular SOD activity detectable in ~10,000 cells per well for typical cell lines |
| Specificity | The xanthine/xanthine oxidase superoxide generation system is specific for superoxide radical under the optimized buffer conditions (pH 8.0, 37°C); cyanide-sensitive Cu/Zn-SOD and cyanide-resistant Mn-SOD can be differentiated by pre-incubating samples with 1–2 mM potassium cyanide; catalase is often added to the reaction to remove H2O2 that could otherwise reduce the rate of the xanthine oxidase reaction |
| Reaction Conditions / Protocol | Prepare cell or tissue lysates in cold PBS with 1% Triton X-100; centrifuge at 10,000 × g for 10 minutes at 4°C; dilute supernatant as needed in dilution buffer; prepare Working Solution (xanthine, WST-1, and buffer); add 20 μL sample or SOD standard to microplate wells; add 200 μL Working Solution; add 20 μL Xanthine Oxidase Solution to initiate formation; immediately begin kinetic reading at 450 nm, taking measurements every 1–2 minutes for 20 minutes at 37°C; calculate SOD activity from the inhibition rate relative to the no-SOD blank |
| Components / Formulation | Xanthine Solution (lyophilized), WST-1 Solution (tetrazolium salt), Xanthine Oxidase (lyophilized, high purity), SOD Standard (purified bovine erythrocyte Cu/Zn-SOD, lyophilized), Assay Buffer (Tris-HCl, pH 8.0, with EDTA), detailed protocol including calculation formula for inhibition rate |
| Storage Conditions | All components at -20°C protected from light; reconstituted xanthine and WST-1 solutions stable 2 weeks at 2–8°C; Xanthine Oxidase solution stable 1 week at 2–8°C |
| Shelf Life | 12 months from date of manufacture |
| Package Specifications | 200 assays, 500 assays (96-well format) |
| Product Form | Lyophilized enzymes and substrate reagents with liquid assay buffer |
| Quality Control | Each lot standardized against reference SOD enzyme preparation; linear inhibition curve (R2 ≥0.97) for the SOD standard dilution series; IC50 for WST-1 reduction within specified range for positive control |
| Key Features | Kinetic format eliminates endpoint timing errors; WST-1-based detection produces water-soluble formazan without extraction steps; high-throughput 96-well compatibility; cyanide differentiation protocol identifies Cu/Zn-SOD versus Mn-SOD contributions to total activity |
For research use only, not for clinical use.
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