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Monitoring the Targeting of Cathepsin D to the Lysosome by Metabolic Labeling and Pulse-chase Analysis.

Authors :
Tavares LA
daSilva LLP
Source :
Bio-protocol [Bio Protoc] 2017 Nov 05; Vol. 7 (21), pp. e2598. Date of Electronic Publication: 2017 Nov 05 (Print Publication: 2017).
Publication Year :
2017

Abstract

Mannose 6-phosphate receptors function can be studied in living cells by investigating alterations in processing and secretion of their ligand Cathepsin D. The assay described here is well established in the literature and comprises the metabolic labeling of newly synthesized proteins with [ <superscript>35</superscript> S] methionine-cysteine in HeLa cells to monitor Cathepsin D processing through secretory pathway and secretion using immunoprecipitation, SDS-PAGE and fluorography.<br /> (Copyright © 2017 The Authors; exclusive licensee Bio-protocol LLC.)

Details

Language :
English
ISSN :
2331-8325
Volume :
7
Issue :
21
Database :
MEDLINE
Journal :
Bio-protocol
Publication Type :
Academic Journal
Accession number :
34595275
Full Text :
https://doi.org/10.21769/BioProtoc.2598