磷酸化 RIPK3 在 L929 细胞、鼠卵黄囊、盲肠和小肠中的免疫染色。
Immunostaining of phospho-RIPK3 in L929 cells, murine yolk sacs, ceca, and small intestines.
机构信息
State Key Laboratory of Cellular Stress Biology, Innovation Center for Cell Biology, School of Life Sciences, Xiamen University, Xiamen, Fujian 361102, China.
State Key Laboratory of Cellular Stress Biology, Innovation Center for Cell Biology, School of Life Sciences, Xiamen University, Xiamen, Fujian 361102, China; Laboratory Animal Center, Xiamen University, Xiamen, Fujian 361102, China; Research Unit of Cellular Stress of CAMS, Cancer Research Center of Xiamen University, Xiang'an Hospital of Xiamen University, School of Medicine, Xiamen University, Xiamen, Fujian 361102, China.
出版信息
STAR Protoc. 2022 Sep 16;3(3):101517. doi: 10.1016/j.xpro.2022.101517. Epub 2022 Jul 1.
Though phospho-receptor-interacting protein 3 (RIP3 or RIPK3) antibodies are used in western blot, immunostaining of murine phospho-RIPK3 is challenging. Here, we verify and describe a detailed protocol for immunofluorescent detection of phospho-RIPK3 in L929 cells and mouse yolk sacs. We also describe in detail the model construction methods, sample preparation steps, and staining procedures for immunohistochemical labeling of RIPK3 activation in mouse ceca and small intestines by utilizing a specific commercially available antibody. For complete details on the use and execution of this protocol, please refer to Zhang et al. (2021) and Wang et al. (2020).
虽然磷酸受体相互作用蛋白 3（RIP3 或 RIPK3）抗体可用于 Western blot，但对鼠磷酸化 RIPK3 的免疫染色具有挑战性。在此，我们验证并描述了一种用于检测 L929 细胞和鼠卵黄囊中磷酸化 RIPK3 的免疫荧光检测的详细方案。我们还详细描述了利用特定的商业可得抗体对鼠盲肠和小肠中 RIPK3 激活的免疫组织化学标记的模型构建方法、样品制备步骤和染色程序。有关此方案使用和执行的完整详细信息，请参阅 Zhang 等人（2021 年）和 Wang 等人（2020 年）。
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