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

3,105 Views

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08:52 min

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April 06, 2022

DOI:

10.3791/63835-v

DOI:
10.3791/63835-v

•

08:52 min
April 06, 2022

•

11 Views
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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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Protein TurnoverSenescent CellsNon-dividing CellsMetabolic LabelingMass SpectrometrySILACProtein HomeostasisCellular StressAgingNeurodegenerative DiseaseLC-MSTrypsinUreaAmmonium BicarbonateDithiothreitolIodoacetamide

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Payea, M., Gorospe, M., Basisty, N. Measurement of Protein Turnover Rates in Senescent and Non-Dividing Cultured Cells with Metabolic Labeling and Mass Spectrometry. J. Vis. Exp. (182), e63835, doi:10.3791/63835 (2022).

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