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Free keywords:
Animals
Antibodies
Cells, Cultured
Fibroblasts/*metabolism
Gene Expression Regulation/physiology
Hippocampus/cytology
Mice
Neurons/metabolism
Proteins/*chemistry/*metabolism
Rats
Staining and Labeling
Abstract:
Protein synthesis is a dynamic process that tunes the cellular proteome in response to internal and external demands. Metabolic labeling approaches identify the general proteomic response but cannot visualize specific newly synthesized proteins within cells. Here we describe a technique that couples noncanonical amino acid tagging or puromycylation with the proximity ligation assay to visualize specific newly synthesized proteins and monitor their origin, redistribution and turnover in situ.