Protocol for the identification and expression analysis of a cytoplasmic membrane-localized protein STING.

STAR Protoc

Department of Phase 1 Trials Center, Harbin Medical University Cancer Hospital, No.150 Haping Road, Nangang District, Harbin, Heilongjiang 150081, China; Key Laboratory of Molecular Oncology of Heilongjiang Province, No.150 Haping Road, Nangang District, Harbin, Heilongjiang 150081, China; Department of Gastrointestinal Medical Oncology, Harbin Medical University Cancer Hospital, No.150 Haping Road, Nangang District, Harbin, Heilongjiang 150081, China. Electronic address:

Published: March 2023

Here, we present a protocol for the detection of the two STING isoforms (erSTING and pmSTING) in human peripheral blood mononuclear cells or mouse splenocytes using Western blot and PCR. We detail steps to construct plasmids encoding each isoform and transfer them into mouse and human cell lines. Finally, we describe how to detect cell membrane localization of pmSTING using flow cytometry, immunoprecipitation, and immunofluorescence. This protocol is applicable for proteins with well-predicted topological structures. For complete details on the use and execution of this protocol, please refer to Li et al..

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10041549PMC
http://dx.doi.org/10.1016/j.xpro.2023.102172DOI Listing

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