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| Product Name | Annexin V-FITC/PI Apoptosis Detection Kit |
| Catalog No. | RESK-0140 |
| Description | This kit uses FITC-labeled Annexin V as a probe to detect the onset of early apoptosis. 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+). The kit is suitable for detection by flow cytometry and fluorescence microscopy. |
| Intended Use | For the detection and discrimination of viable, early apoptotic, late apoptotic and necrotic cells in cell samples. 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 fluorescence microscopy (FITC channel for Annexin V; PI channel for membrane-compromised cells) |
| Sample Type | Cultured cells (adherent or suspension); also applicable to plant cells or bacteria after protoplast preparation; not suitable for tissue sections (frozen or paraffin) |
| Performance Range / Specifications | Convenient 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-FITC and PI in 1x Binding Buffer (containing Ca2+) according to the kit manual, then analyze promptly by flow cytometry or fluorescence microscopy. Result interpretation: viable cells (Annexin V-/PI-); early apoptotic cells (Annexin V+/PI-); late apoptotic and necrotic cells (Annexin V+/PI+); bare nuclei (Annexin V-/PI+). |
| Components / Formulation | Annexin V-FITC conjugate Propidium Iodide (PI), 20 ug/mL Binding Buffer (Ca2+-containing; detailed formulation is proprietary) |
| Storage Conditions | Store at -20°C protected from light; 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 | 5 T 20 T 50 T 100 T |
| Product Form | Liquid kit (ready-to-use fluorescent probes and buffer) |
| Concentration | PI: 20 ug/mL |
| Molecular Weight | Annexin V: 35-36 kDa |
| pH Range / Optimal pH | Binding Buffer at pH 7.5 (substitute buffers must be supplemented with 2.5 mM CaCl2 and adjusted to pH 7.5) |
| 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 microscopes with FITC (Ex/Em approximately 488/525 nm) and PI detection channels |
| Recommended Buffer System | 1x Binding Buffer supplied in the kit; if a substitute is needed, use PBS or Hanks' buffer supplemented with 2.5 mM CaCl2, adjusted to pH 7.5 |
| Application Notes / Precautions | Annexin V detects early apoptosis, whereas JC-1 detects mid-stage apoptosis and TUNEL detects late-stage apoptosis. For special samples such as anucleate red blood cells, or when drugs such as doxorubicin interfere with PI detection, Annexin V-FITC may be used alone without PI. The kit cannot be used to stain tissue sections (frozen or paraffin-embedded). For plant cells or bacteria, protoplasts must be prepared first; the staining volume only needs to cover the cells, and staining time varies among cell types. |
For research use only, not for clinical use.
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