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| Product Name | Annexin V-Fluor 488/PI Apoptosis Detection Kit |
| Catalog No. | RESK-0143 |
| Description | This kit uses Annexin V labeled with Fluor 488, a green fluorescent dye, as a probe to detect the onset of early apoptosis. Detection can be performed by flow cytometry or other fluorescence-based instruments. In viable cells, phosphatidylserine (PS) is localized on the inner leaflet of the plasma membrane. During early apoptosis, PS is translocated from the inner to the outer leaflet and becomes exposed to the extracellular environment. Annexin V is a Ca2+-dependent phospholipid-binding protein with a molecular weight of 35-36 kDa that binds PS with high affinity, allowing fluorescently labeled Annexin V to bind to the membrane of early apoptotic cells via the exposed PS. Propidium iodide (PI) is a nucleic acid dye that cannot penetrate the intact membrane of viable or early apoptotic cells, but can enter late apoptotic and necrotic cells and stain the nucleus red. When Annexin V and PI are used in combination, PI is excluded from viable cells (Annexin V-/PI-) and early apoptotic cells (Annexin V+/PI-), while late apoptotic and necrotic cells are co-stained and appear double positive (Annexin V+/PI+). |
| Intended Use | For the detection and discrimination of viable, early apoptotic, late apoptotic and necrotic cells by flow cytometry or fluorescence-based instruments. For research use only. |
| Principle / Technology | In viable cells, phosphatidylserine (PS) is localized on the inner leaflet of the plasma membrane. During early apoptosis, PS is translocated from the inner to the outer leaflet and becomes exposed to the extracellular environment. Annexin V is a Ca2+-dependent phospholipid-binding protein with a molecular weight of 35-36 kDa that binds PS with high affinity, allowing fluorescently labeled Annexin V to bind to the membrane of early apoptotic cells via the exposed PS. Propidium iodide (PI) is a nucleic acid dye that cannot penetrate the intact membrane of viable or early apoptotic cells, but can enter late apoptotic and necrotic cells and stain the nucleus red. When Annexin V and PI are used in combination, PI is excluded from viable cells (Annexin V-/PI-) and early apoptotic cells (Annexin V+/PI-), while late apoptotic and necrotic cells are co-stained and appear double positive (Annexin V+/PI+). |
| Detection Method | Flow cytometry or other fluorescence detection instruments (green fluorescence channel for Annexin V-Fluor 488; PI channel for membrane-compromised cells) |
| Sample Type | Cultured cells (adherent or suspension) |
| Performance Range / Specifications | Convenient operation and accurate detection |
| Specificity | Annexin V binds externalized phosphatidylserine in a Ca2+-dependent manner with high affinity; PI is excluded by intact membranes and stains only late apoptotic/necrotic cells |
| Reaction Conditions / Protocol | Stain cells with Annexin V-Fluor 488 and PI in 1x Binding Buffer according to the kit manual, then analyze by flow cytometry or other fluorescence detection instruments. Viable cells (Annexin V-/PI-); early apoptotic cells (Annexin V+/PI-); late apoptotic/necrotic cells (Annexin V+/PI+). |
| Components / Formulation | Annexin V-Fluor 488 conjugate Propidium Iodide (PI) Binding Buffer (Ca2+-containing) |
| Storage Conditions | Store at -20°C protected from light for long-term storage; avoid repeated freeze-thaw cycles. For repeated use within a short period, store at 4°C protected from light. |
| Shelf Life | 1 year when stored at -20°C protected from light; 6 months when stored at 4°C protected from light |
| Package Specifications | 20 T 50 T 100 T |
| Product Form | Liquid kit (ready-to-use fluorescent probes and buffer) |
| Molecular Weight | Annexin V: 35-36 kDa |
| Shipping Conditions | Shipped on wet ice |
| Expiration Date / Stability | Stable for 1 year at -20°C protected from light (avoid repeated freeze-thaw); stable for 6 months at 4°C protected from light |
| Regulatory / Compliance | For research use only |
| Compatibility | Flow cytometers and fluorescence detection instruments with a green (FITC-type) detection channel and a PI channel |
| Recommended Buffer System | 1x Binding Buffer supplied in the kit |
| Application Notes / Precautions | Annexin V detects early apoptosis, whereas JC-1 detects mid-stage apoptosis and TUNEL detects late-stage apoptosis. Protect the fluorescent reagents from light during handling and incubation. |
For research use only, not for clinical use.
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