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Biochemistry

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JoVE Journal Biochemistry
Measurement of Protein Turnover Rates in Senescent and Non-Dividing Cultured Cells with Metabolic Labeling and Mass Spectrometry
 

Measurement of Protein Turnover Rates in Senescent and Non-Dividing Cultured Cells with Metabolic Labeling and Mass Spectrometry

Article DOI: 10.3791/63835-v • 08:52 min • April 6th, 2022
April 6th, 2022 •
1, 1, 2
1Laboratory of Genetics and Genomics, National Institute on Aging Intramural Research Program, National Institutes of Health, 2Translational Gerontology Branch, National Institute on Aging Intramural Research Program, National Institutes of Health

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This protocol describes the workflow for metabolic labeling of senescent and non-dividing cells with pulsed SILAC, untargeted mass spectrometry analysis, and a streamlined calculation of protein half-lives.

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Keywords: Protein Turnover Senescent Cells Non-dividing Cells Metabolic Labeling Mass Spectrometry SILAC Protein Homeostasis Cellular Stress Aging Neurodegenerative Disease LC-MS Trypsin Urea Ammonium Bicarbonate Dithiothreitol Iodoacetamide
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