| Product Name | Annexin V-mCherry/SYTOX Green Apoptosis Detection Reagent |
| Catalog No. | CCAT-HMM-0027 |
| Description | Apoptosis is distinguished from necrosis by characteristic morphological and biochemical changes, including cell shrinkage, nuclear shrinkage, and fragmentation of the nuclear membrane and nucleolus. In normal living cells, phosphatidylserine (PS) is located on the cytoplasmic side of the cell membrane. In apoptotic cells, PS is internalized from the inner side of the plasma membrane to the cell surface (the outer side of the cell membrane), thereby exposing PS to the extracellular environment. During the apoptosis of white blood cells, the PS marker on the surface of white blood cells can be recognized by macrophages, ultimately leading to their phagocytosis by macrophages. Human vascular anticoagulant Annexin V is a 35-36 kDa Ca²⁺-dependent phospholipid-binding protein with high affinity for PS. Cells stained with Annexin V labeled with the red fluorescent protein mCherry (Annexin V-mCherry) can be easily and directly detected using fluorescence microscopy, laser confocal microscopy, or flow cytometry, which are fluorescence detection devices, to identify the outward flipping of phosphatidylserine, a key feature of apoptosis. It should be noted that in necrotic cells, due to the disruption of cell membrane integrity, PS located on the inner side of the cell membrane is also stained by Annexin V-mCherry. SYTOX Green is a non-cell membrane-permeable fluorophore that cannot penetrate the bioactive cytoplasmic membrane but can pass through the disorganized regions of dead cell membranes to reach the nucleus, where it binds to DNA double helices to form SYTOX Green-DNA complexes, resulting in a fluorescence intensity increase of over 500 times and producing bright green fluorescence. Therefore, SYTOX Green can only stain dead cells, making it useful for distinguishing normal cells from necrotic cells. |
| Application | This product uses the apoptosis fluorescent probe Annexin V-mCherry in combination with the dead cell green fluorescent probe SYTOX Green to detect apoptosis and necrosis. |
| Spectral Parameters | Annexin V-mCherry: Ex/Em (nm) = 587/610; SYTOX Green: Ex/Em (nm) = 504/523 |
| Applicable Instruments | Flow cytometer, fluorescence microscope, or other fluorescence detection equipment |
| Materials to Bring | 1 × PBS, pH 7.2-7.6; sterile ddH₂O |
| Storage | Store at -20°C away from light. |
| Shelf Life | 1 year |
| Notes | If there is bacterial or fungal contamination, it will severely affect the detection results. After staining, testing should be conducted as soon as possible. Prolonged waiting time may lead to an increase in the number of apoptotic or necrotic cells. If trypsin was used during cell collection, care must be taken to remove any residual trypsin. Residual trypsin will digest and degrade Annexin V-mCherry, ultimately resulting in staining failure. Fluorescent substances are prone to quenching. When performing fluorescence observations, minimize observation time and ensure light-protected storage during handling and storage. For your safety and health, please wear a lab coat and disposable gloves during operation. |
For research use only, not for clinical use.
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