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| Product Name | RealTime-Glo MT Cell Viability Assay, Luciferase-Based |
| Catalog No. | CATR-HMM-0054 |
| Description | A homogeneous, non-lytic, bioluminescent cell viability assay that continuously measures the reducing potential of living cells in real time. Based on a genetically engineered thermostable luciferase and a cell-permeable pro-substrate, this assay provides real-time kinetic viability data over 72 hours without the need to terminate the culture or add additional reagents after the initial addition. The glow-type luminescence signal is stable for hours, enabling batch plate processing. |
| Intended Use | Real-time continuous cell viability monitoring; drug mechanism-of-action studies with temporal resolution; identification of optimal compound treatment windows; long-term cytotoxicity profiling (24–72 hours); co-culture and 3D model viability assessment. |
| Principle / Technology | Cell-permeable pro-substrate enters viable cells and is reduced by intracellular reducing environment; reduced substrate diffuses into culture medium where it is utilized by luciferase to generate stable luminescence; signal is proportional to the number of metabolically active cells at each time point |
| Detection Method | Bioluminescence; luminescence plate reader; integration time 0.5–1.0 seconds per well |
| Sample Type | Adherent and suspension mammalian cells; compatible with 3D spheroids and co-cultures; primary cells and immortalized cell lines |
| Sensitivity / LOD | Detects as few as 10 cells per well; linear range 10–50,000 cells (96-well); signal stable >4 hours per reading window |
| Specificity | Specific for viable, metabolically active cells; signal diminishes rapidly upon cell death as reducing environment collapses; no interference from phenol red, serum, or common media supplements |
| Reaction Conditions / Protocol | Add reagent at 2× concentration in culture medium at time of plating or treatment; equilibrate 1 hour at 37 °C; read luminescence at desired time points; incubate between readings at 37 °C, 5% CO2 |
| Components / Formulation | NanoLuc luciferase enzyme, cell-permeable MT Cell Viability Substrate, assay buffer; all provided as 1,000× concentrate |
| Storage Conditions | Store at -80 °C for long-term; -20 °C for up to 3 months; protect from light |
| Shelf Life | 6 months at -20 °C; 12 months at -80 °C from manufacture date |
| Package Specifications | 10 mL (100 assays), 50 mL (500 assays), 100 mL (1,000 assays) |
| Product Form | Concentrated enzyme and substrate solutions in proprietary buffer |
| Key Features | Real-time non-lytic measurement — monitor same wells repeatedly over 72 hours; 72-hour reagent stability at 37 °C; glow-type signal (half-life >5 hours) enables batch processing; add-once-and-read format — no additional reagent additions; compatible with HTS automation; Z' factor >0.7 for screening applications |
| Purity | NanoLuc luciferase specific activity >10^10 RLU/mg; endotoxin <0.01 EU/µg protein |
| Concentration | As specified on product label; working concentrations optimized per protocol |
| Activity / Unit Definition | Signal-to-noise ratio and linearity verified on reference cell lines per lot |
| Molecular Weight | Varies by dye or reagent component as specified |
| Source / Origin | Synthetic dyes and tetrazolium compounds; recombinant enzymes where applicable |
| pH Range / Optimal pH | pH 7.2–7.4 for cell-based assays |
| Shipping Conditions | Dry ice -80 °C; do not allow to thaw during transit |
| Expiration Date / Stability | 12 months at -80 °C; 6 months at -20 °C; thaw on ice before use; prepare single-use aliquots to avoid freeze-thaw cycles; working solution stable 24 hours at 4 °C or 8 hours at room temperature |
| Regulatory / Compliance | For laboratory and research use only; RUO; manufactured under ISO 9001; not classified as dangerous goods; contains no hazardous components at use concentration |
| Compatibility | Compatible with all standard cell culture media formulations including those with phenol red; serum concentrations up to 20% do not interfere; compatible with antibiotics (penicillin-streptomycin, gentamicin); test DMSO at >0.5% as carrier solvent may affect viability independently |
| Recommended Buffer System | PBS or phenol red-free complete medium as specified in protocol |
| Application Notes / Precautions | For optimal temporal resolution, read luminescence at 1–4 hour intervals. Include cell-free medium + reagent background control and 100% kill control (e.g., 0.1% Triton X-100 or 70% ethanol treatment) to define baseline. For drug treatment studies, add reagent 1 hour before first reading to allow equilibrium. Normalize data to time-zero readings for fold-change viability plots. |
| Batch-to-Batch Consistency | Signal linearity R² ≥0.99 with reference cell number standard curve per lot |
For research use only, not for clinical use.
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