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

Abstract

Summary: 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.1. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• Cell Biology
• Cell isolation
• Flow Cytometry/Mass Cytometry
• Cell Membrane
• Cancer
• Sequencing