Lakshmanan Imayavaramban, Batra Surinder K
Biochemistry and Molecular Biology, Nebraska Medical Center, Omaha, USA.
Bio Protoc. 2013 Mar 20;3(6). doi: 10.21769/bioprotoc.374.
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.
该检测方法用于计数发生凋亡的细胞数量。通过先用膜联蛋白V和碘化丙啶溶液对细胞进行染色,然后进行流式细胞术分析来检测凋亡。其原理基于正常细胞本质上具有疏水性,因为它们在内膜(面向细胞质的一侧)表达磷脂酰丝氨酸,而当细胞发生凋亡时,内膜翻转成为外膜,从而使磷脂酰丝氨酸暴露。膜联蛋白V可检测暴露的磷脂酰丝氨酸,碘化丙啶可对坏死细胞进行染色,坏死细胞的DNA含量有泄漏,这有助于区分凋亡细胞和坏死细胞。