Elabscience® Annexin V-AF488 / PI Cell Apoptosis Detection kit is developed to identify apoptotic and necrotic cells.
Annexin V is a member of the annexin family, which binds to phosphatidylserine (PS) in a calcium-dependent manner. Annexin V-AF488, the AF488-conjugated format, binds specifically to the PS on the outer leaflet of apoptotic cell membrane and can be detected with flow cytometry or fluorescence microscopy.
Propidium Iodide (PI) is a common DNA dye that is not permeable to cell membrane. Once binding to DNA, the flurescence of PI increases by nearly 20 fold. Due to the loss of integrity of membrane, PI can enter late apoptotic or necrotic cells to stain DNA. Cells at different apoptotic stages can be distinguished by using Annexin V and PI.
|E-CK-A137||Annexin V-AF488 Reagent||100 μL||250 μL||500 μL||1 mL||2~8°C|
|E-CK-A151||Annexin V Binding Buffer (10 ×)||1.4 mL×2||5.5 mL||11 mL||11 mL×2||2~8°C|
|E-CK-A161||Propidium Iodide (PI) Staining Solution (50μg/mL)||100 μL||250 μL||500 μL||1 mL||2~8°C|
Jurkat cells were treated with 5 μM Camptothecin and detected with this kit.
Jurkat cells were cultured with (Left) or without (Right) 5 μM Camptothecin for 4 h. Annexin V-AF488 single-positive cells were early apoptotic cells, Annexin V-AF488 and PI double-positive cells were necrotic or late apoptotic cells, and PI single-positive cells were naked nuclei.
1.For maximal assay performance, this kit should be used within 12 months. Avoid freeze / thaw cycles.
2.For FCM analysis, please set untreated cells stained with both Annexin V-AF488 and PI as negative control. As for compensation controls, please use drug-treated cells stained with either Annexin V-AF488 or PI.
3.Annexin V-AF488 can be detected in FITC channel while PerCP/Cy5.5 channel is preferred to ECD channel fo.
4.Detect apoptosis as soon as possible after staining to avoid the increase number of apoptosis or necrosis.
5.Avoid extended exposure of the samples to direct light to protect the fluorophores from quenching.
6.For your safety and health, please wear the lab coat and disposable gloves before the experiments.
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