Annexin v-apc conjugate (C09-1097-438)

Product introduction：Annexin V (annexin-V) is a ca2+ - dependent phospholipid binding protein with a molecular weight of 35-36 KD, which can selectively bind to phosphatidylserine (PS). Phosphatidylserine (PS) is mainly distributed on the inner side of the cell membrane, that is, the side adjacent to the cytoplasm. At the early stage of apoptosis, different types of cells will evert phosphatidylserine to the cell surface and expose it to the extracellular environment. At this time, annexin V labeled with fluorescent protein APC, namely annexin v-apc, is combined with everted phosphatidylserine (PS), and the everted phosphatidylserine, an important feature of apoptosis, can be directly detected by flow cytometry. Normal cells will not be stained by annexin v-apc, and apoptotic or necrotic cells will be stained by annexin v-apc. Annexin v-apc can be combined with some non permeable nuclear dyes (7-aad/pi) to distinguish cells at different stages of apoptosis.Product parameters：Annexin v-apc:ex/em = 650/660 NMMatters needing attention：1. please centrifuge the product to the bottom of the tube immediately before use, and then conduct subsequent experiments. 2. to reduce the process of apoptosis, the incubation process can be operated on ice, but the incubation time should be extended to at least 30 min. 3. as apoptosis is a rapid process, it is recommended that samples be analyzed within 1 h after staining. 4. for adherent cells, digestion is a key step. If there are floating cells when adherent cells induce apoptosis, the floating cells and adherent cells should be collected and stained. Handle adherent cells with care to avoid artificial damage to cells. The trypsin digestion time is too short, and the cells need to be blown hard to fall off, which is easy to cause damage to the cell membrane and excessive PI intake; If the digestion time is too long, the cell membrane is also prone to damage, and even affect the binding of phosphatidylserine and annexin v-apc on the cell membrane. When digesting, spread pancreatin on the bottom of the well plate, fully contact the pancreatin with the cells when shaking gently, then pour out most of the pancreatin, use the remaining small amount of pancreatin to digest for a period of time, and terminate when the gap between cells increases and the bottom of the bottle is spotted. Try not to use EDTA in the digestive juice, which will affect the binding of annexin V to PS. 5. after the adherent cells are digested with trypsin, it is recommended to stain after recovering in the optimal culture conditions and medium for about 30 min to avoid false positives. 6. in order to avoid losing cells when washing cells, you can use a large tip over a small tip to aspirate. 7. the optimal concentration of dye is determined by the specific experimental requirements. 8. fluorescent dyes have quenching problems. Please try to avoid light during storage and use to slow down fluorescence quenching. 9. for your safety and health, please wear experimental clothes and disposable gloves.Scope of application：Early apoptosis detection, annexin v-coupled fluorescent protein.