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Annexin V/PI apoptosis kit
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产品名称:
Annexin V/PI apoptosis kit
产品型号:
AP101-30
产品展商:
上海博蕴生物科技有限公司
简单介绍
Introduction: Annexin V/PI apoptosis kitAnnexin V Apoptosis Detection Kit is based on the observation that soon after initiating apoptosis, cells translocate the membrane Annexin V/PI
Annexin V/PI apoptosis kit的详细介绍
III. Annexin V-FITC Assay Protocol: Annexin V/PI apoptosis kit
A. Incubation of cells with Annexin V-FITC Annexin V/PI apoptosis kit
1. Induce apoptosis by desired method. Annexin V/PI apoptosis kit
2. Collect 1-5 x 105 cells by centrifugation. Annexin V/PI apoptosis kit
3 Prepare 1X annexin-binding buffer. For example, for ~10 assays, add 1 mL of 5X annexin-binding buffer to 4 mL of deionized water
4. Resuspend cells in 500 μl of 1X Binding Buffer. Annexin V/PI apoptosis kit
5. Add 5 μl of Annexin V-FITC and 10 μl of propidium iodide. Annexin V/PI apoptosis kit
6. Incubate at room temperature for 5 min in the dark. Annexin V/PI apoptosis kit
Proceed to B or C below depending on method of analysis. Annexin V/PI apoptosis kit
B. Quantification by Flow Cytometry Annexin V/PI apoptosis kit
Analyze Annexin V-FITC binding by flow cytometry (Ex = 488 nm; Em = 530 nm) using FITC signal detector (usually FL1) and PI staining by the phycoerythrin emission signal detector (usually FL2). Annexin V/PI apoptosis kit
For adherent cells, gently trypsinize and wash cells once with serum-containing media before incubation with Annexin V-FITC (A.3-5).
C. Detection by Fluorescence Microscopy Annexin V/PI apoptosis kit
1. Place the cell suspension from Step A.5 on a glass slide. Cover the cells with a glass coverslip. Annexin V/PI apoptosis kit
For analyzing adherent cells, grow cells directly on a coverslip. Following incubation (A.5), invert coverslip on a glass slide and visualize cells. The cells can also be washed and fixed in 2% formaldehyde before visualization. (Cells must be incubated with Annexin V-FITC before fixation since any cell membrane disruption can cause nonspecific binding of Annexin V to PS on the inner surface of the cell membrane.) Annexin V/PI apoptosis kit
2. Observe the cells under a fluorescence microscope using a dual filter set for FITC & rhodamine.
Cells that have bound Annexin V-FITC will show green staining in the plasma membrane. Cells that have lost membrane integrity will show red staining (PI) throughout the nucleus and a halo of green staining (FITC) on the cell surface (plasma membrane).
Note: These products are distributed from K101-400, which is from biovison Research Products Annexin V/PI apoptosis kit