Detection of membrane-anchoring lipid modifications of proteins in cells by radioactive metabolic labeling.
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.
Palabras clave
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