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| Product Name | Calcein-AM / Propidium Iodide Live-Dead Cell Double Staining Kit |
| Catalog No. | CATR-HMM-0039 |
| Description | A two-color fluorescence staining system for simultaneous discrimination of live and dead cells. Calcein-AM (acetoxymethyl ester) passively enters cells and is hydrolyzed by intracellular esterases in live cells to produce green fluorescent calcein, while propidium iodide penetrates only damaged membranes of dead cells, staining nuclei red. |
| Intended Use | Rapid visual or quantitative assessment of cell viability in mixed cell populations for cytotoxicity testing, tissue engineering quality control, and flow cytometry-based viability analysis. |
| Principle / Technology | Non-fluorescent Calcein-AM crosses intact cell membranes and is cleaved by ubiquitous intracellular esterases to membrane-impermeable green-fluorescent calcein (Ex/Em: 495/515 nm); propidium iodide enters only cells with compromised membranes, intercalating into DNA and emitting red fluorescence (Ex/Em: 535/617 nm). |
| Detection Method | Fluorescence microscopy with FITC and TRITC filter sets, or flow cytometry with 488 nm excitation |
| Sample Type | Adherent and suspension mammalian cells, primary cultures, stem cell-derived cells, tissue fragments; suitable for fluorescence microscopy and flow cytometry applications |
| Performance Range / Specifications | Stains 100–1,000,000 cells per assay; calcein retention time >2 hours in viable cells at 37°C |
| Sensitivity / LOD | Detection of single live and dead cells distinguishable by both microscopy and flow cytometry |
| Specificity | Calcein-AM activation requires functional intracellular esterase activity; PI exclusively labels nuclei of membrane-compromised cells; minimal cross-fluorescence between the two channels with appropriate filter sets |
| Reaction Conditions / Protocol | Prepare cell suspension or adherent culture; wash with PBS or assay buffer; add staining solution containing 2 μM Calcein-AM and 4 μM PI; incubate 15–30 minutes at 37°C; wash cells; analyze by fluorescence microscopy (FITC/TRITC) or flow cytometry (FL1/FL2 channels) |
| Components / Formulation | Calcein-AM solution (1 mg/mL in anhydrous DMSO), Propidium Iodide solution (1 mg/mL in water), 10× Assay Buffer (PBS-based), detailed staining protocol |
| Storage Conditions | -20°C protected from light and moisture; Calcein-AM stock sensitive to hydrolysis; prepare working solution fresh before each use; PI stable at -20°C for 12 months |
| Shelf Life | 12 months from date of manufacture for individual stock solutions |
| Package Specifications | 200 assays, 500 assays, 1,000 assays (counting chambers or flow cytometry format) |
| Product Form | Individual concentrated stock solutions in DMSO (Calcein-AM) and aqueous buffer (PI), with separate assay buffer |
| Quality Control | Each lot tested on control live and ethanol-killed cells to verify proper differential staining; Calcein-AM hydrolysis efficiency confirmed by enzymatic activity; PI membrane exclusion validated on live cells |
| Key Features | Rapid 15-minute staining protocol; compatible with fluorescence microscopy, flow cytometry, and plate reader formats; can be multiplexed with additional fluorophores; discriminates live from dead cells in heterogeneous populations |
For research use only, not for clinical use.
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