Journal
/
Biologie
/
Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry
Journal JoVE
Biologie
This content is Free Access.
Journal JoVE Biologie
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

•

06:09 min

•

July 31, 2011

•

, , , , , , , ,
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

Chapitres

  • 00:05Titre
  • 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

Traduction automatique

Traduction automatique

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

Article

Embed

AJOUTER À LA PLAYLIST

Usage Statistics

Vidéos Connexes

MALDI Sample Preparation: the Ultra Thin Layer Method

A Quantitative Assessment of The Yeast Lipidome using Electrospray Ionization Mass Spectrometry

Analyzing Large Protein Complexes by Structural Mass Spectrometry

In vivo Quantification of G Protein Coupled Receptor Interactions using Spectrally Resolved Two-photon Microscopy

Profiling of Methyltransferases and Other S-adenosyl-L-homocysteine-binding Proteins by Capture Compound Mass Spectrometry (CCMS)

Profilering Thiol Redox Proteome Brug Isotope Tagging massespektrometri

Identifikation af protein-komplekser i<em> Escherichia coli</em> Hjælp Sekventiel Peptide Affinitetsrensning i kombination med Tandem massespektrometri

Capture Compound massespektrometri - Et kraftfuldt værktøj til at identificere nye C-di-GMP effektorproteiner

Valgt Reaktion Overvågning massespektrometri for Absolute Protein Kvantificering

Berigelse af ekstracellulære matrixproteiner fra væv og fordøjelse til peptider for massespektrometri Analyse

Read Article