Apoptosis refers to the autonomous and orderly death of cells controlled by genes in order to maintain the stability of the internal environment. Apoptosis is different from cell necrosis, cell apoptosis generally refers to a programmed cell death process in which cells are regulated by intracellular genes and their products during development or under the action of some factors. Cell necrosis is a process of cell death caused by strong physical, chemical or biological factors. The difference between apoptosis and necrosis lies in the characteristic morphological and biochemical changes, including changes in cell membrane permeability and nuclear chromatin, cytoplasmic contraction and loss of membrane asymmetry.
The detection kit is a double fluorescence kit for detecting apoptosis based on Oxazoleyellow and PI dyes. This kit is suitable for fluorescence microscope, flow cytometry, luciferase labeling and other fluorescence detection systems.
Oxazoleyellow is a kind of carbonyl cyanine monomer green fluorescent dye with non-cell membrane penetration and high affinity for DNA. It basically has no fluorescence when it does not bind to DNA, but it can emit bright green fluorescence when it binds to DNA. When apoptosis occurs, the membrane permeability changes, and Oxazoleyellow can enter the cell to bind to DNA and emit bright green fluorescence, so it is often used to detect apoptosis. It should be noted that Oxazoleyellow can also stain dead cells, so double staining with dead cell-specific fluorescent staining of PI is needed to effectively determine apoptosis.
PI (Propidium Iodide) is a red fluorescent dye that can dye DNA. It is an analogue of pyridine bromide, which emits red fluorescence after intercalation of double-stranded DNA. Although PI cannot pass through the living cell membrane, it can dye the nucleus through the damaged cell membrane of dead cells. Therefore, the combination of Oxazoleyellow and PI can be directly used to detect apoptosis. Apoptotic cells show green fluorescence, dead cells show both red and green fluorescence positive, living cells have little or no fluorescence.
Note:
1. Please centrifuge the product to the bottom of the tube immediately before use, and then carry out the follow-up experiment.
2. All fluorescent dyes have the problem of quenching, please try to avoid light in order to slow down fluorescence quenching.
3. For your safety and health, please wear a lab coat and disposable gloves.
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