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Updated: Aug 23, 2026
Simultaneous Visualization of the Dynamics of Crosslinked and Single Microtubules In Vitro by TIRF Microscopy
Published on: February 18, 2022
Protocol for systematic identification of microtubule luminal proteins using knock-in BioID proximity labeling and
Zhengrong Zhou1, Jinhui Shao2, Weitong Li3
1Joint Shantou International Eye Center of Shantou University and The Chinese University of Hong Kong, Shantou, Guangdong 515041, China; Neuroscience center, Department of Basic Medical Sciences, Shantou University Medical College, Shantou, Guangdong 515041, China; State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
Abstract:
Systematic elucidation of microtubule (MT) luminal proteins provides insights into MT biology and its molecular regulation. Here, we present a protocol for screening and validating these proteins using proximity labeling, mass spectrometry, fluorescence imaging and western blot (WB). We describe steps for constructing a biotin ligase knock-in cell line and performing BioID screening. We detail how to perform fluorescence imaging and WB to validate the MT luminal proteins. This protocol enables systematic identification to advance research on MT luminal proteins. For complete details on the use and execution of this protocol, please refer to Shao et al.1.
