Annexin V-EGFP PI апоптосис клетки комплект для обнаружения антител

Apoptosis is a normal physiological process that occurs in the process of embryonic development and maintenance of tissue homeostasis. It is accompanied by many morphological changes. Among them, the loss of cell membrane is one of the early characteristics of apoptosis. In normal cells, phosphotidylserine (PS) is only distributed on the inner side of the phospholipid bilayer of the cell membrane. However, in the early stage of apoptosis, PS will flip from the inner side of the lipid membrane to the outer side, exposing it to the outside of the cell. Annexin V (Annexin V) is a Ca2+-dependent phospholipid binding protein that has a high affinity for PS and specifically binds to cells exposed to PS. Therefore, Annexin V is used as one of the indicators for detecting early cell apoptosis. Propidium Iodide (PI) is a nucleic acid dye. It cannot penetrate normal cells with intact cell membranes and early apoptotic cells, but can penetrate the cell membranes of late apoptotic and necrotic cells and stain the nucleus.

This product uses EGFP (enhanced Green Fluorescent Protein) and Annexin V to form a fusion protein as a detection probe to detect early cell apoptosis. At the same time, PI is used to distinguish surviving cells from necrotic and late apoptotic cells. When Annexin V-EGFP is used in combination with PI, living cells show negative staining (Annexin V-/PI-), early apoptotic cells show single fluorescence positive (Annexin V+/PI-), and late apoptotic cells and necrotic cells show dual fluorescence Positive (Annexin V+/PI+). This kit is suitable for flow cytometer or fluorescence microscope detection. At the same time, because EGFP and Annexin V are fused 1:1, this product can also quantitatively detect apoptotic cells.

Component:

1. During the entire experiment, the operation should be as gentle as possible to avoid cell breakage and affect the experimental results.

2. Washing the cells with PBS cannot be omitted, and at the same time, remove the residual PBS as much as possible.

3. When using trypsin to digest cells, you should be careful to avoid artificial damage to the cells and control the digestion time. If the digestion time is too short, the cells need to be blown off to fall off, which is likely to cause mechanical damage to the cell membrane; if the digestion time is too long, the cell membrane is also vulnerable to damage. Affect the test results. In addition, pancreatin containing EDTA cannot be used. EDTA will affect the binding of Annexin V and PS.

4. After the adherent cells are stimulated by apoptosis, if some of the cells float, it is necessary to collect the cell culture supernatant and the adherent cells and stain them at the same time, which will make the results more accurate.

5. Annexin V-EGFP and PI are sensitive to light, please avoid light during operation. The test should be carried out as soon as possible after the reaction.

6. Please wear laboratory clothes and disposable gloves during the experiment to avoid contamination and ensure safety.

This product is for scientific research purposes only, not for clinical diagnosis!