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Immunopeptidomics: Isolation of Mouse and Human MHC Class I- and II-Associated Peptides for Mass Spectrometry Analysis
JoVE Journal
Biochemistry
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JoVE Journal Biochemistry
Immunopeptidomics: Isolation of Mouse and Human MHC Class I- and II-Associated Peptides for Mass Spectrometry Analysis

Immunopeptidomics: Isolation of Mouse and Human MHC Class I- and II-Associated Peptides for Mass Spectrometry Analysis

DOI:
10.3791/63052-v

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09:32 min

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October 15, 2021

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, , , ,
1CHU Sainte-Justine Research Center, 2Department of Pathology and Cellular Biology, Faculty of Medicine,Université de Montréal

Chapters

  • 00:04Introduction
  • 00:41Coupling Antibodies to Sepharose CNBr-Activated Beads
  • 02:57Cell Lysis and Immunocapture with Antibody Coupled-Beads
  • 04:06Elution of MHC Peptides
  • 07:11Results: Analysis of the MHC-Binding Affinity
  • 08:56Conclusion

Summary

Automatic Translation

Automatic Translation

Here, we present a protocol for the purification of MHC class I and class II peptide complexes from mouse and human cell lines providing high-quality immunopeptidomics data. The protocol focuses on sample preparation using commercially available antibodies.

Tags

Keywords: ImmunopeptidomicsMHC Class IMHC Class IIPeptide IsolationMass SpectrometryHumanMouseAutoimmune DiseasesCancerVirologySepharose BeadsAntibody CouplingBlockingWashingCell LysisPeptide Elution

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