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Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification
JoVE Journal
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JoVE Journal Biyokimya
Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification

Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification

11,716 Views

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10:37 min

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November 15, 2017

DOI:

10.3791/56474-v

DOI:
10.3791/56474-v

•

10:37 min
November 15, 2017

•

7 Views
, , , , , , ,
1St. Jude Proteomics Facility,St. Jude Children’s Research Hospital, 2Departments of Structural Biology and Developmental Neurobiology,St. Jude Children’s Research Hospital, 3Heilongjiang University of Chinese Medicine

Özet

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We present a protocol to accurately quantitate proteins with isobaric labelling, extensive fractionation, bioinformatics tools, and quality control steps in combination with liquid chromatography interfaced to a high-resolution mass spectrometer.

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Proteome ProfilingIsobaric LabelingLiquid ChromatographyMass SpectrometryQuantificationProtein IdentificationProtein QuantitationBiomarker DiscoveryCell LysisProtein DigestionAcetonitrileLys-C ProteaseDTTHEPES

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High, A. A., Tan, H., Pagala, V. R., Niu, M., Cho, J., Wang, X., Bai, B., Peng, J. Deep Proteome Profiling by Isobaric Labeling, Extensive Liquid Chromatography, Mass Spectrometry, and Software-assisted Quantification. J. Vis. Exp. (129), e56474, doi:10.3791/56474 (2017).

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