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Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry
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Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry

Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry

DOI:
10.3791/2812-v

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06:09 min

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July 31, 2011

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, , , , , , , ,
1Clinical Research Division,Fred Hutchinson Cancer Research Center – FHCRC, 2Department of Biochemistry and Microbiology,University of Victoria, 3Broad Institute of MIT and Harvard, 4Genome BC Proteomics Centre,University of Victoria, 5Plasma Proteome Institute

Chapters

  • 00:05Title
  • 01:05Trypsin Enzymatic Digestion and Cleanup
  • 03:17Peptide Immunoaffinity Enrichment
  • 04:45Analysis by Multiple Reaction Monitoring – Mass Spectrometry
  • 05:24MRM of a SISCAPA-Enriched Sample
  • 05:44Conclusion

Summary

Automatic Translation

Automatic Translation

Stable Isotope Standards and Capture by Anti-Peptide Antibodies (SISCAPA) couples affinity enrichment of peptides with stable isotope dilution mass spectrometry (MRM-MS) to provide quantitative measurement of peptides as surrogates for their respective proteins. Here we describe the protocol using magnetic particles in a partially automated format.

Tags

QuantificationProteinsPeptide Immunoaffinity EnrichmentMass SpectrometryTraditional ImmunoassaysSISCAPAStable Isotope StandardsCapture By Anti-peptide AntibodiesSRM/MRM-MSSmall MoleculesProtein ConcentrationProteotypic PeptidesTrypsin

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