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| Product Name | Annexin V-FITC / Propidium Iodide Apoptosis Detection Flow Cytometry Kit |
| Catalog No. | CCAT-HMM-0043 |
| Description | A dual-parameter fluorescence kit that discriminates early apoptotic, late apoptotic/necrotic, and viable cells by simultaneously detecting phosphatidylserine externalization with FITC-conjugated Annexin V and membrane integrity with propidium iodide. The combined staining pattern reveals the mode and progression of cell death. |
| Intended Use | Quantitative detection of apoptosis versus necrosis in drug mechanism-of-action studies, toxicology, immunology (CTL-mediated killing), and evaluation of cancer chemotherapeutic agent efficacy. |
| Principle / Technology | In viable cells, phosphatidylserine is actively maintained on the inner leaflet of the plasma membrane by flippase enzymes; during early apoptosis, phosphatidylserine translocates to the outer leaflet where it is bound by calcium-dependent Annexin V-FITC; propidium iodide enters only late apoptotic and necrotic cells with compromised membrane integrity; dual labeling produces three distinct populations: Annexin V-/PI- (viable), Annexin V+/PI- (early apoptotic), Annexin V+/PI+ (late apoptotic/necrotic). |
| Detection Method | Flow cytometry with 488 nm excitation; FITC fluorescence in FL1 channel (530/30 nm), PI fluorescence in FL2/PE channel (575/26 nm); also compatible with fluorescence microscopy |
| Sample Type | Freshly harvested single-cell suspensions from adherent or suspension mammalian cell cultures; dissociated primary tissue cells; blood lymphocytes after density gradient separation |
| Performance Range / Specifications | Resolves ≥1% apoptotic cells in heterogeneous populations; staining specificity >95% for early apoptosis in well-characterized inducers (staurosporine, anti-Fas, etoposide) |
| Sensitivity / LOD | Detection of early apoptotic cells comprising approximately 2–5% of total event count with clear separation from viable populations |
| Specificity | Annexin V binding to phosphatidylserine is calcium-dependent and reversible by EDTA chelation; PI staining is specific for membrane-compromised cells; necrotic cells display Annexin V+/PI+ pattern in the absence of calcium chelator |
| Reaction Conditions / Protocol | Harvest cells (2–5 × 10^5 per sample), wash twice with cold PBS, resuspend in 100 μL 1× Binding Buffer; add 5 μL Annexin V-FITC and 5 μL PI; incubate 15 minutes at room temperature in the dark; add 400 μL 1× Binding Buffer; analyze by flow cytometry within 1 hour; include unstained, Annexin V-only, and PI-only single-color compensation controls |
| Components / Formulation | Annexin V-FITC conjugate (in Tris-based buffer with BSA and sodium azide), Propidium Iodide solution, 10× Binding Buffer (HEPES buffered saline with CaCl2), detailed protocol with gating strategy and compensation matrix recommendations |
| Storage Conditions | 2–8°C protected from light for all components; do not freeze; stable for 12 months |
| Shelf Life | 12 months from date of manufacture |
| Package Specifications | 50 tests, 100 tests, 200 tests (flow cytometry format) |
| Product Form | Liquid reagents including fluorescent conjugates in stabilizing buffer |
| Quality Control | Each lot validated using Jurkat cells treated with camptothecin or staurosporine as positive apoptosis controls; compensation matrices confirmed; lot-to-lot consistency in mean fluorescence intensity within 20% |
| Key Features | Simultaneous discrimination of cell death modes; rapid 15-minute staining protocol; FITC conjugate compatible with all benchtop flow cytometers; single-color controls included for compensation setup; widely published and accepted methodology for apoptosis research |
For research use only, not for clinical use.
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