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| Product Name | XTT Cell Viability Assay Kit (sodium 3'-[1-(phenylaminocarbonyl)-3,4-tetrazolium]-bis (4-methoxy-6-nitro) benzene sulfonic acid) |
| Catalog No. | CATR-HMM-0047 |
| Description | A tetrazolium-based colorimetric assay kit using XTT reagent that is reduced to a water-soluble orange formazan product by metabolically active cells in conjunction with an electron coupling reagent. The water-soluble formazan avoids the dissolution step needed in MTT assays, enabling a homogeneous format. |
| Intended Use | Cell proliferation and viability measurement in high-throughput compound library screening and large-scale cytotoxicity profiling of anticancer drug candidates. |
| Principle / Technology | XTT (sodium 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide inner salt) is reduced at the cell surface by mitochondrial succinate dehydrogenase via trans-plasma membrane electron transport, with PMS (N-methyl dibenzopyrazine methyl sulfate) serving as an intermediate electron carrier, producing a soluble formazan absorbing at 450 nm. |
| Detection Method | Colorimetric absorbance at 450 nm with reference wavelength at 650 nm using a microplate reader |
| Sample Type | Adherent and suspension mammalian cells in 96-well or 384-well microplates; also suitable for yeast and filamentous fungi viability assessment |
| Performance Range / Specifications | Linear: 500–100,000 cells per well depending on cell type; incubation 2–5 hours; assay sensitivity and signal intensity are typically lower than WST-1 for equivalent cell numbers |
| Sensitivity / LOD | Approximately 1,000–2,000 cells per well for common vertebrate cell types |
| Specificity | XTT reduction is electron transport chain-dependent and specific to metabolically active cells; background absorbance in cell-free medium is very low; PMS reagent can be toxic to cells at extended incubations beyond 4 hours |
| Reaction Conditions / Protocol | Prepare XTT/PMS working solution (mix 5 mL XTT reagent with 0.1 mL PMS activator); add 50 μL per well to 100–200 μL culture volume; incubate 2–5 hours at 37°C with 5% CO2; shake briefly; read absorbance at 450 nm against 650 nm reference; no medium removal needed |
| Components / Formulation | XTT sodium salt (lyophilized or 1 mg/mL solution), PMS (phenazine methosulfate) electron coupling reagent, sterile PBS, instruction manual with cell type optimization guidelines |
| Storage Conditions | XTT: -20°C protected from light; PMS: -20°C protected from light and moisture; XTT/PMS working solution fresh use only |
| Shelf Life | 12 months at -20°C for lyophilized reagent |
| Package Specifications | 1,000 tests, 5,000 tests (96-well format) |
| Product Form | Lyophilized XTT with separate liquid PMS activator |
| Quality Control | Each lot tested for crosstalk (minimum 0.1 OD difference between medium-only and maximum cell density wells); linearity R2 ≥0.98 against known cell standards; sterility verified |
| Key Features | Homogeneous add-and-read protocol eliminates organic solvent step present in MTT protocol; water-soluble formazan product; amenable to kinetic reading for time-course experiments during the incubation window |
For research use only, not for clinical use.
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