Detection of membrane-anchoring lipid modifications of proteins in cells by radioactive metabolic labeling.
| Autor: | 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. |
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| Jazyk: | angličtina |
| Zdroj: | STAR protocols [STAR Protoc] 2023 Sep 15; Vol. 4 (3), pp. 102416. Date of Electronic Publication: 2023 Jul 04. |
| DOI: | 10.1016/j.xpro.2023.102416 |
| Abstrakt: | 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 . Competing Interests: Declaration of interests The authors declare no competing interests. (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.) |
| Databáze: | MEDLINE |
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