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Detection of membrane-anchoring lipid modifications of proteins in cells by radioactive metabolic labeling.
Bhat, Sameer Ahmed; Ferguson, Rachel; Adhikari, Ritika; Kuchay, Shafi.
  • Bhat SA; Department of Biochemistry and Molecular Genetics, University of Illinois at Chicago, MBRB #1252, Chicago, IL 60607, USA.
  • Ferguson R; Department of Biochemistry and Molecular Genetics, University of Illinois at Chicago, MBRB #1252, Chicago, IL 60607, USA.
  • Adhikari R; Department of Biochemistry and Molecular Genetics, University of Illinois at Chicago, MBRB #1252, Chicago, IL 60607, USA.
  • Kuchay S; Department of Biochemistry and Molecular Genetics, University of Illinois at Chicago, MBRB #1252, Chicago, IL 60607, USA. Electronic address: kuchay@uic.edu.
STAR Protoc ; 4(3): 102416, 2023 Sep 15.
Article en En | MEDLINE | ID: mdl-37405928
ABSTRACT
Prenylation and palmitoylation are two major lipid modifications of cellular proteins that anchor proteins to cell membranes. Here, we present a protocol for detecting these modifications in cellular proteins by radioactive metabolic labeling. We describe steps for metabolic labeling of cells, cell harvesting for carrying out immunoprecipitations, subjecting immunocomplexes to SDS-PAGE, and transferring them to polyvinylidine flouride (PVDF) membranes. We then detail detection of labeled target proteins by exposing PVDF membranes to phosphor screens and using a phosphor imager machine. For complete details of this protocol, please refer to Liang et al.1.
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Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Polímeros de Fluorocarbono / Proteínas / Lípidos de la Membrana Tipo de estudio: Diagnostic_studies Idioma: En Año: 2023 Tipo del documento: Article
Texto completo
Imprimir
XML
PubMed Links
Search on Google

Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Polímeros de Fluorocarbono / Proteínas / Lípidos de la Membrana Tipo de estudio: Diagnostic_studies Idioma: En Año: 2023 Tipo del documento: Article