YEASEN은 생명과학 연구 및 진단을 위한 고품질의 연구용 시약과 기기를 제공하는 글로벌 바이오 기업입니다.
제품 설명
Annexin V-YSFluorTM 647/PI Apoptosis Detection Kit
제품 번호
40304ES20 / size: 20T
40304ES50 / size: 50T
40304ES60 / size: 100T
제품 설명
Product name | Cat# | Specification |
Annexin V-YSFluorTM 647/PI Apoptosis Detection Kit | 40304ES20 | 20 T |
| 40304ES50 | 50 T |
| 40304ES60 | 100 T |
Annexin V-YSFluorTM 647/PI Apoptosis Detection Kit is an Annexin Fluor 647 labeled with Annexin V as a probe to detect the occurrence of early apoptosis, which can be detected by flow cytometry or other fluorescence detection equipment.
The detection principle is that in normal living cells, phosphotidylserine (PS) is located on the inner side of the cell membrane, but in early apoptotic cells, PS reverses from the inner side to the surface of the cell membrane and is exposed to the extracellular environment. Annexin V is a Ca2+ -dependent phospholipid binding protein with a molecular weight of 35-36 kD that binds PS with high affinity. Phosphatidylserine can bind to the membrane of early-apoptotic cells by exposing it laterally.
In addition, Propidium Iodide (PI) is provided in this kit to distinguish between surviving early cells and necrotic or late apoptotic cells. PI is a kind of nucleic acid dye, which can not penetrate the intact cell membrane of normal cells or early apoptotic cells, but can penetrate the cell membrane of late apoptotic and necrotic cells and make the cell nucleus red. Therefore, when Annexin V was used in combination with PI, PI was excluded from living cells (Annexin V-/PI-) and early apoptotic cells (Annexin V+/PI-). The apoptotic and necrotic cells were double positive by YSFluorTM 647 and PI binding staining (Annexin V+/PI+).This kit requires flow cytometry.
Component |
| 40304ES20（20T） | 40304ES50（50T） | 40304ES60（100T） |
40304-A | Annexin V-YSFluorTM 647 | 100 μL | 250 μL | 500 μL |
40304-B | PI Staining Solution (20 μg/mL) | 200 μL | 500 μL | 1.0 mL |
40304-C | 1×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.
1. Sample dyeing
1.1 suspension cells: 300 g, centrifuged at 4℃ for 5 mins to collect cells.
Adherent cells: After digestion with trypsin without EDTA, cells were collected by centrifugation at 300 g at 4℃ for 5 mins. Trypsin digestion time should not be too long to prevent false positive.
1.2. Cells were washed twice with pre-cooled PBS, 300 g each time, centrifuged at 4℃ for 5 mins.
1.3. Discard PBS and add 100 μL 1×Binding Buffer to resuscitate cells.
1.4. Add 5 μL Annexin V-YSFluorTM 647 and 10 μL PI, mix gently.
1.5. The reaction was conducted at room temperature for 15 mins, away from light.
1.6. 400 μL 1×Binding Buffer was added, and the samples were mixed and placed on ice. The samples were detected by flow cytometry or fluorescence microscope within 1 hour.
【Notes】: In order to avoid the loss of cells when washing cells, a small tip can be used to suck liquid.
2. Flow cytometry analysis
YSFluorTM 647 has a maximum excitation wavelength of 651 nm and a maximum emission wavelength of 667 nm; The maximum excitation wavelength of pi-DNA complex is 535 nm, and the maximum emission wavelength is 615 nm. Analysis was performed using CellQuest and other software to plot two-color dot plots with YSFluorTM647 as abscissa and PI as ordinates. 10,000 events were collected for each sample.
YEASEN의 모든 제품을 만나 보세요!
YEASEN - Official Distributor in South Korea "Morebio" 한국 공식 대리점 "모아바이오"
MSDS
온라인 견적