BD Pharmingen™ Propidium Iodide Staining Solution

Flow cytometric analysis of cells following induction of apoptosis. Jurkat leukemia cells were left untreated (left top & left bottom panels) or treated for 5 hours (middle top & middle bottom panels) or 12 hours (right top & right bottom panels) with anti-human Fas antibody (clone DX2, Cat. No. 555670) and Protein G (the addition of Protein G enhances the ability of DX2 to induce apoptosis, presumably by cross-linking Fas). Cells were incubated with Annexin V-Biotin, (Cat. No. 556417) followed by incubation with SAv-FITC (Cat. No. 554060) in Propidium Iodide (PI) Staining Solution (Cat. No. 556463). Cells were then analyzed by flow cytometry. Untreated cells were primarily Annexin V-Biotin and PI negative, indicating that they were viable and not undergoing apoptosis. After a 5 hour treatment with DX2, there were two populations of cells: Cells undergoing apoptosis (Annexin V-Biotin positive and PI negative), and cells that were viable and not undergoing apoptosis (Annexin V-Biotin and PI negative). After a 12 hour treatment with DX2, three populations of cells were identified: Cells that had already died or were in late stage of apoptosis (Annexin V-Biotin and PI positive), cells undergoing apoptosis (Annexin V-Biotin positive and PI negative), and cells that were viable and not undergoing apoptosis (Annexin V-Biotin and PI negative).

Flow cytometric analysis of cells following induction of apoptosis. Jurkat leukemia cells were left untreated (left top & left bottom panels) or treated for 5 hours (middle top & middle bottom panels) or 12 hours (right top & right bottom panels) with anti-human Fas antibody (clone DX2, Cat. No. 555670) and Protein G (the addition of Protein G enhances the ability of DX2 to induce apoptosis, presumably by cross-linking Fas). Cells were incubated with Annexin V-Biotin, (Cat. No. 556417) followed by incubation with SAv-FITC (Cat. No. 554060) in Propidium Iodide (PI) Staining Solution (Cat. No. 556463). Cells were then analyzed by flow cytometry. Untreated cells were primarily Annexin V-Biotin and PI negative, indicating that they were viable and not undergoing apoptosis. After a 5 hour treatment with DX2, there were two populations of cells: Cells undergoing apoptosis (Annexin V-Biotin positive and PI negative), and cells that were viable and not undergoing apoptosis (Annexin V-Biotin and PI negative). After a 12 hour treatment with DX2, three populations of cells were identified: Cells that had already died or were in late stage of apoptosis (Annexin V-Biotin and PI positive), cells undergoing apoptosis (Annexin V-Biotin positive and PI negative), and cells that were viable and not undergoing apoptosis (Annexin V-Biotin and PI negative).