Annexin 5 fitc assay kit

Experimental samples were collected, washed twice with 1xPBS, re-suspended in binding buffer, and stained using an Annexin V-FITC assay Kit (Fisher Scientific, Pittsburgh, PA). The cells were incubated with 50 μg/ml Annexin V and 10 μg/ml propidium iodide in 1x binding buffer at room temperature in the dark, for 15 min, and immediately analyzed by flow cytometry (EPICS XL). Annexin V-positive/PI-negative cells were considered to be early apoptotic, cells with both Annexin V and PI positive to be late apoptotic, and Annexin V negative but PI positive to be “necrotic” [11 (link)]. Percent of cell death was calculated as the sum of the percentages of both apoptotic and necrotic cells.

Following the indicated exposure of either HL60 cells or ex vivo AML blasts to AraC followed by D2/CA combination, the cells were collected, washed twice with 1xPBS, then re-suspended in the binding buffer, and stained using an Annexin V-FITC assay Kit (Fisher Scientific, Pittsburgh, PA). The cells were incubated with 50 μg/ml Annexin V and 10 μg/ml propidium iodide in 1x binding buffer at room temperature in the dark for 15 min, and immediately analyzed by flow cytometry (EPICS XL). Annexin V-positive/PI-negative cells were considered to be early apoptotic, cells with both Annexin V and PI positive to be late apoptotic, and Annexin V negative but PI positive to be “necrotic”(18 (link)).