免疫荧光染色法检测原代培养的小鼠海马神经元中的磷酯酰肌醇。
Immunofluorescence staining of phosphoinositides in primary mouse hippocampal neurons in dissociated culture.
机构信息
State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Innovation Academy for Seed Design, Chinese Academy of Sciences, Beijing 100101, China.
State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Innovation Academy for Seed Design, Chinese Academy of Sciences, Beijing 100101, China; College of Life Sciences, University of Chinese Academy of Sciences, Beijing 100039, China.
出版信息
STAR Protoc. 2022 Sep 16;3(3):101549. doi: 10.1016/j.xpro.2022.101549. Epub 2022 Jul 16.
Phosphoinositides (PIPs) are low-abundant membrane lipids with critically important functions in cellular physiology. To investigate their subcellular distribution in neurons, we optimized protocols for immunofluorescence staining of intracellular or plasma membrane PIPs with commercially available antibodies. Here, we describe the preparation and transfection of primary mouse hippocampal neurons in dissociated culture, followed by immunofluorescence staining and quantitative analysis of PIP signals. In addition, we expand the application of the protocol to proteins located at the cytoplasmic leaflet of cellular membranes. For complete details on the use and execution of this protocol, please refer to Guo et al. (2022).
磷酯酰肌醇（PIPs）是一种低丰度的膜脂，在细胞生理学中具有至关重要的功能。为了研究其在神经元中的亚细胞分布，我们使用商业上可获得的抗体优化了用于检测细胞内或质膜 PIPs 的免疫荧光染色方案。在这里，我们描述了在分离培养中制备和转染原代小鼠海马神经元的过程，随后进行 PIP 信号的免疫荧光染色和定量分析。此外，我们还将该方案的应用扩展到位于细胞膜细胞质小叶的蛋白质。有关该方案使用和执行的完整详细信息，请参见 Guo 等人（2022 年）。
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