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| Product Name | Mitochondrial Complex II (Succinate Dehydrogenase) Activity Microplate Assay Kit |
| Catalog No. | CMTR-HMM-0074 |
| Description | Colorimetric assay for mitochondrial Complex II (succinate dehydrogenase, SDH) activity in isolated mitochondria, tissue homogenates, and permeabilized cells. Uses DCIP as artificial electron acceptor whose reduction is monitored at 600 nm. Inhibitors (azide, antimycin A) block Complex III/IV to eliminate electron backflow, ensuring specific Complex II activity measurement. |
| Intended Use | Mitochondrial Complex II activity measurement; respiratory chain function assessment; mitochondrial disorder research; Complex II inhibitor/activator screening; drug mitochondrial toxicity evaluation. |
| Principle / Technology | Succinate dehydrogenase oxidizes succinate to fumarate; decylubiquinone serves as electron acceptor; coupled DCIP reduction (blue to colorless) monitored at 600 nm |
| Detection Method | Colorimetric; kinetic at 600 nm; absorbance decrease over 10-30 min |
| Sample Type | Isolated mitochondria (10-50 µg protein), tissue homogenates, permeabilized cells |
| Sensitivity / LOD | Detects activity from 5 µg mitochondrial protein; Km (succinate) ~0.3 mM |
| Specificity | Specific for Complex II; azide/antimycin A block Complex III/IV; malonate included for specificity validation |
| Reaction Conditions / Protocol | Add mitochondria to plate with succinate, DCIP, decylubiquinone, azide, antimycin A; read kinetically at 600 nm |
| Components / Formulation | Assay buffer (10×), succinate (1 M), DCIP (4 mM), decylubiquinone (2.5 mM), sodium azide (100 mM), antimycin A (1 mg/mL), malonate inhibitor control (500 mM) |
| Storage Conditions | Azide/antimycin A at -20 °C; DCIP at 4 °C protected from light; decylubiquinone at -20 °C |
| Shelf Life | 6 months from manufacture date |
| Package Specifications | 100 assays in 96-well format |
| Product Form | Liquid reagents with DMSO/ethanol stock solutions |
| Key Features | Direct Complex II measurement bypassing Complex III/IV; malonate inhibitor for validation; DCIP at 600 nm avoids mitochondrial turbidity; optimized for kinetic plate readers |
| Purity | DCIP >97%; decylubiquinone >98% |
| Concentration | As specified per kit format; probe stock solutions in DMSO as specified |
| Activity / Unit Definition | Standard curve linearity R² ≥0.99 per lot; dynamic range as specified |
| Molecular Weight | Varies by probe and buffer component |
| Source / Origin | Synthetic fluorescent probes; recombinant enzymes; purified metabolite standards |
| pH Range / Optimal pH | pH 7.2–7.4 for cell-based assays; pH varies for biochemical metabolite detection |
| Shipping Conditions | Cold pack -20 °C; DCIP at 4 °C; ship within 48 hr |
| Expiration Date / Stability | 6 months; DCIP light-sensitive — discard on color change; decylubiquinone 1 month at -20 °C |
| Regulatory / Compliance | For laboratory and research use only; RUO; sodium azide toxic; antimycin A respiratory toxin; ISO 9001 |
| Compatibility | Compatible with fluorescence microplate readers, luminometers, and spectrophotometers |
| Recommended Buffer System | Assay-specific buffer optimized per product protocol |
| Application Notes / Precautions | Normalize signal to cell number or protein content; avoid probe photobleaching; include appropriate antioxidant controls for ROS assays |
| Batch-to-Batch Consistency | Standard curve parameters within ±15% of reference lot |
For research use only, not for clinical use.
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