Protocol for systematic identification of microtubule luminal proteins using knock-in BioID proximity labeling and fluorescence imaging validation
Year: 2026
Type: article
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
Source: STAR Protocols
Institutions: Chinese University of Hong Kong, Chinese Academy of Sciences, Shantou University, Shantou University Medical College, Institute of Genetics and Developmental Biology, State Key Laboratory of Molecular Developmental Biology, Joint Shantou International Eye Center, Capital Medical University, Capital Institute of Pediatrics, University of Chinese Academy of Sciences
Language: English
Topic: Biotin and Related Studies
Subfield: Cell Biology
Domain: Life Sciences
Open Access status: gold
Estimated APC paid: $1,550
OpenAlex ID: W7203900897