| Background | Apoptosis is a gradually orchestrated process of biochemical reactions of a cell from an organism. It can be triggered by many stimuli, including infection, hypoxia, ischemia, nutrient removal, toxins, heat, radiation, drugs, chemicals, and disease. Consequently, these stresses alter the morphology of a cell, including cell shrinkage, nuclear and cytoplasmic condensation, chromatin fragmentation, membrane blebbing, and apoptotic body formation. CHAMOT fluorescent dye (iFluor 488) conjugated Annexin V is highly purified product. During early apoptosis, cells will translocate membrane phosphatidylserine (PS) from the inner face of the membrane to the cell surface. Propidium iodide (PI) is a common fluorescent dye to detect DNA. It can be used in flow cytometry to evaluate the cell cycle and cell viability during apoptosis. The product can be used in one-step staining procedure without wash step within 20 minutes. |
| Stability & Storage | 2-8°C for 12 months under sterile conditions from date of receipt. |
| Shipping | Blue Ice |
| Components | CM001-50D | CM001-100D |
|---|---|---|
|
Annexin V-iFluor 488 |
1 × 0.25 mL |
1 × 0.5 mL |
|
PI Solution |
1 × 0.25 mL |
1 × 0.5 mL |
|
10×Binding Buffer |
2 × 2 mL |
3 × 2 mL |
Procedure
(1) Collect 1-5x10^5 cells in the flow tube by centrifugation.
(2) Wash cells in 2 mL cold phosphate-buffered saline (PBS) and collect by centrifugation.
(3) Re-suspend cell in 500 μL of 1X Binding Buffer.
(4) Add 5 μL of Annexin V-iFluor 488 and 5 μL of PI, and gently mix the cells and incubate for 15-20 minutes at RT in the dark.
(5) After incubation, the samples should be kept on ice and perform flow cytometry using filters appropriated for fluorescein (FITC, corresponding to Annexin V-iFluor 488)
No experimental data is available
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