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Måling av proteinomsetningshastigheter i senescent og ikke-dele kultivert celler med metabolsk merking og massespektrometri
JoVE Journal
Biochemistry
This content is Free Access.
JoVE Journal Biochemistry
Measurement of Protein Turnover Rates in Senescent and Non-Dividing Cultured Cells with Metabolic Labeling and Mass Spectrometry
DOI:

08:52 min

April 06, 2022

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Chapters

  • 00:05Introduction
  • 00:58Labeling of Cells Pulsed SILAC and Harvest of Lysates
  • 02:29In-Solution Trypsin Digestion and Sample Clean-Up with Solid-Phase Extraction (SPE)
  • 04:15Data-Dependent Acquisition Mass Spectrometry Analysis and Calculation of Protein Half-Lives
  • 06:31Results: Measurement of Protein Turnover Rates in Senescent and Non-Dividing Cultured Cells Using pSILAC Mass Spectrometry
  • 08:13Conclusion

Summary

Automatic Translation

Denne protokollen beskriver arbeidsflyten for metabolsk merking av senescent og ikke-delende celler med pulserende SILAC, umålt massespektrometrianalyse og en strømlinjeformet beregning av proteinhalvvanser.

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