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Bottom-up og Shotgun Proteomics at identificere en omfattende Cochlear Proteome
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Biologia
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JoVE Journal Biologia
Bottom-up and Shotgun Proteomics to Identify a Comprehensive Cochlear Proteome

Bottom-up og Shotgun Proteomics at identificere en omfattende Cochlear Proteome

DOI:
10.3791/51186-v

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14:23 min

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March 07, 2014

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,
1Department of Otolaryngology, Morsani College of Medicine,University of South Florida

Capitoli

  • 00:05Titolo
  • 02:13Protein Extraction
  • 03:03Endoproteinase LysC and Tryptic Protein Digestion of Whole Lysate Using Filter Assisted Sample Preparation
  • 05:34Ion Exchange Chromatography
  • 06:20GELFREE Fractionation of Cochlear Sensory Epithelium
  • 07:54Protein Digestion of GELFREE Fractions Using Filter Assisted Sample Preparation
  • 10:07Results: Identification of membrane and Soluble Proteins by High-resolution Mass Spectrometry Analysis
  • 14:00Conclusion

Summary

Traduzione automatica

Traduzione automatica

Proteomanalyse af cochlear sensoriske epithel kan være en udfordring på grund af sin lille størrelse, og fordi membranproteiner er vanskeligt at isolere og identificere. Både membranen og opløselige proteiner, kan identificeres ved at kombinere flere præparative metoder og separationsteknikker sammen med høj opløsning massespektrometri.

Tags

Keywords: ProteomicsCochlear ProteomeMass SpectrometryProtein IdentificationSeparation TechniquesSample Fractionation2D-DIGEAntibody MicroarraySCXRPGELFrEE

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