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| Product Name | Cell Counting Kit-F (Fluorometric, Resazurin-Based) |
| Catalog No. | CATR-HMM-0052 |
| Description | A fluorometric cell viability and proliferation assay kit based on the reduction of resazurin (Alamar Blue indicator) to highly fluorescent resorufin by metabolically active cells. The non-toxic nature of the reagent allows for continuous monitoring of the same cell population over time, making it ideal for kinetic proliferation studies, drug sensitivity time-course experiments, and long-term cytotoxicity assessment. The fluorescent readout (Ex/Em = 560/590 nm) provides superior sensitivity compared to colorimetric methods. |
| Intended Use | Fluorometric quantification of viable cell number; kinetic cell proliferation monitoring; drug sensitivity time-course assays; cytotoxicity screening; cell health assessment in 3D spheroid and organoid cultures. |
| Principle / Technology | Metabolically active cells reduce non-fluorescent resazurin (blue) to highly fluorescent resorufin (pink) via NAD(P)H-dependent reductase enzymes; fluorescence intensity is proportional to viable cell number |
| Detection Method | Fluorometric; excitation 530–570 nm, emission 580–610 nm; also readable by absorbance at 570 nm with reference at 600 nm |
| Sample Type | Adherent and suspension mammalian cells; compatible with 3D spheroid and organoid cultures; primary cells and established cell lines |
| Sensitivity / LOD | Detects as few as 50–100 cells per well (96-well format); linear range 100–50,000 cells depending on cell type |
| Reaction Conditions / Protocol | Add reagent at 10% (v/v) of culture medium volume; incubate at 37 °C, 5% CO2 for 1–4 hours; read fluorescence; for kinetic studies, measure at multiple time points without washing |
| Components / Formulation | Resazurin sodium salt (Alamar Blue) in sterile PBS, pH 7.4; ready-to-use solution |
| Storage Conditions | Store at 2–8 °C; protect from light; do not freeze |
| Shelf Life | 12 months from manufacture date; use within 6 months after opening |
| Package Specifications | 5 mL (500 tests), 25 mL (2,500 tests), 100 mL (10,000 tests) in amber bottles |
| Product Form | Sterile, ready-to-use dark blue liquid |
| Key Features | Non-toxic — allows continuous monitoring of same cell population; 10× more sensitive than MTT assay; simple add-and-read protocol; no washing, fixing, or solubilization steps; compatible with serum-containing and serum-free media; validated for 96-well, 384-well, and 1536-well formats |
| Purity | Resazurin sodium salt purity ≥95% by HPLC; endotoxin <0.05 EU/mL |
| 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 | Cold pack 2–8 °C; protect from light during transit |
| Expiration Date / Stability | 12 months at 2–8 °C protected from light; 6 months after opening; discard if reagent turns pink prior to use, indicating reduction |
| Regulatory / Compliance | For laboratory and research use only; RUO classification; manufactured under ISO 9001; not classified as dangerous goods |
| Compatibility | Compatible with DMEM, RPMI-1640, MEM, F-12, and most standard cell culture media with or without phenol red; phenol red may increase background — use phenol red-free medium for maximum sensitivity |
| Recommended Buffer System | PBS or phenol red-free complete medium as specified in protocol |
| Application Notes / Precautions | For optimal results, include a cell-free blank (medium + reagent) and serially diluted cell standards to generate a standard curve. Incubation time varies by cell type: rapidly dividing cells (HeLa, HEK293) 1–2 hours; slower cells (primary fibroblasts) 3–4 hours. For 3D cultures, extend incubation to 4–6 hours to allow reagent penetration. Protect plate from light during incubation and reading. |
| 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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