Elabscience® Annexin V-EGFP / DAPI 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-EGFP, the EGFP-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.
4',6-Diamidino-2-Phenylindole (DAPI) has a high DNA binding constant and is efficiently excluded by intact cells. It is useful for DNA analysis and dead cell discrimination during flow cytometric analysis. Due to the loss of integrity of membrane, DAPI can enter late apoptotic or necrotic cells to stain DNA. Cells at different apoptotic stages can be distinguished by using Annexin V and DAPI.
|Cat.||Products||20 Assays||50 Assays||100 Assays||200 Assays||Storage|
|E-CK-A119||Annexin V-EGFP 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-A163||DAPI Reagent||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-EGFP single-positive cells were early apoptotic cells, Annexin V-EGFP and DAPI double-positive cells were necrotic or late apoptotic cells, and DAPI 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-EGFP and DAPI as negative control. As for compensation controls, please use drug-treated cells stained with either Annexin V-EGFP or DAPI.
3.Annexin V-EGFP can be detected in FITC channel while DAPI channel is preferred to Pacific Blue channel.
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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