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Samtidig affinitetberigelse af to ændringer efter oversættelser til kvantificering og lokalisering af lokaliteter på stedet
Journal JoVE
Biologie
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Journal JoVE Biologie
Simultaneous Affinity Enrichment of Two Post-Translational Modifications for Quantification and Site Localization

Samtidig affinitetberigelse af to ændringer efter oversættelser til kvantificering og lokalisering af lokaliteter på stedet

DOI:
10.3791/60780-v

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12:11 min

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February 27, 2020

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, , , , ,
1Buck Institute for Research on Aging

Chapitres

  • 00:04Introduction
  • 01:19Desalting of Non-Enriched Proteolytic Peptides via Large-Scale Solid-Phase Extraction
  • 02:46Simultaneous Enrichment of K-acetylated and K-succinylated Peptides with Immunoaffinity Beads
  • 04:30Elution of the Peptides Bound to Antibody Beads
  • 07:02Data Acquisition Using DDA (Data-Dependent Acquisition) and DIA (Data-Independent Acquisition)
  • 10:02Results: One-pot Enrichment of PTMs
  • 11:19Conclusion

Summary

Traduction automatique

Traduction automatique

Denne arbejdsgang beskriver ydeevnen af tids- og omkostningseffektiv berigelse af flere proteinændringer efter oversættelser (PTM)samtidig til kvantitativ global proteomisk analyse. Protokollen udnytter peptid-niveau PTM berigelse med flere konjugerede antistoffer, efterfulgt af data-uafhængige erhvervelse massespektrometri analyse for at få biologisk indsigt i PTM krydstale.

Tags

Simultaneous Affinity EnrichmentPost-translational ModificationsQuantificationSite LocalizationMass SpectrometryAcetylationSuccinylationProtein ModificationsPTMsReproducibilityC18 ResinImmunoaffinity Purification

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