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제품 설명
Annexin V- YSFluorTM 488/PI Apoptosis Detection Kit
제품 번호
40305ES20 / size: 20T
40305ES50 / size: 50T
40305ES60 / size: 100T
제품 설명
Product name | Cat# | Specification |
Annexin V- YSFluorTM 488/PI Apoptosis Detection Kit
| 40305ES20 | 20 T |
| 40305ES50 | 50 T |
| 40305ES60 | 100 T |
The Annexin V-YSFluorTM 488/PI Apoptosis Detection Kit used YSFluorTM 488-labeled Annexin V as a probe to detect early Apoptosis, which could be detected by flow cytometry or other fluorescence Detection devices.
The detection principle is as follows: in normal living cells, phosphotidylserine (PS) is located in the inner side of the cell membrane, but in early apoptotic cells, PS flips from the inner side of the cell membrane to the surface of the cell membrane, and is exposed to the extracellular environment. Annexin-V is a Ca2+ -dependent phospholipid-binding protein with molecular weight of 35 to 36KD, which binds with high affinity to PS and binds to the cell membrane of early apoptotic cells through phosphatidylserine exposed on the outside of the cell.
In addition, Propidium Iodide (PI) is provided to distinguish surviving early cells from necrotic or late apoptotic cells. PI is a nucleic acid dye, which can not pass through the intact cell membrane of normal cells or early apoptotic cells, but can pass through the cell membrane of late apoptotic and necrotic cells and make the nucleus red. Thus, when Annexin V was combined with PI, PI was excluded from living cells (Annexin V-/PI-) and early apoptotic cells (Annexin V+/PI-). In contrast, late apoptotic and necrotic cells were double positive for YSFluorTM 488 and PI (Annexin V+/PI+).
Component |
| 40305ES20（20T） | 40305ES50（50T） | 40305ES60（100T） |
40305-A | Annexin V-YSFluorTM 488 | 100 μL | 250 μL | 500 μL |
40305-B | PI Staining Solution (20 μg/mL) | 200 μL | 500 μL | 1.0 mL |
40305-C | 4×Binding Buffer | 10 mL | 25 mL | 50 mL |
The components are shipped with ice pack and can be stored at -20°C for 1 year.
b) adherent cell: After digestion with trypsin without EDTA, cells were harvested by centrifugation at 300 g for 5 min at 4 ° C. Trypsin digestion time should not be too long to prevent false positives.
2. The cells were washed twice with PBS precooled at 4 ° C, each time using 300g, and centrifuged at 4 ° C for 5 min.
3. Cells were resuspended by adding 250 μL 1×Binding Buffer, and the concentration was adjusted to 1×106/mL.
4. 100 μL of cell suspension was added into a 5mL flow cytometry tube, 5μL Annexin V-YSFluorTM 488 and 10 μL PI were added, and gently mixed.
5. The reaction was kept away from light and kept at room temperature for 15 min.
6. 400 μL of 1×Binding Buffer was added, mixed, and the samples were detected by flow cytometry within 1 hour.
【Notes】：To avoid cell loss during cell washing, a small Tip can be used to aspirate fluid.
YSFluorTM 488 has a maximum excitation wavelength of 488 nm and a maximum emission wavelength of 519 nm;The maximum excitation wavelength of PI-DNA complex was 535 nm, and the maximum emission wavelength was 615 nm. CellQuest and other software were used to analyze and draw two-color dot plot with YSFluorTM 488 as abscissa and PI as ordinate. Collect 10,000 events per sample.
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