Protocol for Apoptosis Assay by Flow Cytometry Using Annexin V Staining Method

This assay is used to count the number of cells that have undergone apoptosis. Apoptosis will be detected by initially staining the cells with Annexin V and propidium iodide solution followed by flow cytometry analysis. It is based on the principle that normal cells are hydrophobic in nature as they express phosphatidyl serine in the inner membrane (side facing the cytoplasm) and when the cells undergo apoptosis, the inner membrane flips to become the outer membrane, thus exposing phosphatidyl serine. The exposed phosphatidyl serine is detected by Annexin V, and propidium iodide stains the necrotic cells, which have leaky DNA content that help to differentiate the apoptotic and necrotic cells., 该测定用于计数已经历凋亡的细胞的数目。 通过用膜联蛋白V和碘化丙啶溶液初始染色细胞，随后通过流式细胞术分析来检测细胞凋亡。 它是基于这样的原理，正常细胞本质上是疏水的，因为它们在内膜（面向细胞质的一侧）表达磷脂酰丝氨酸，并且当细胞经历细胞凋亡时，内膜翻转成为外膜，从而暴露磷脂酰丝氨酸。 暴露的磷脂酰丝氨酸由膜联蛋白V检测，碘化丙啶染色坏死细胞，其具有有助于区分凋亡和坏死细胞的渗漏DNA含量。