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用于大规模核相互作用分析的无标签免疫沉淀质谱工作流程
JoVE Journal
Biyokimya
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JoVE Journal Biyokimya
Label-Free Immunoprecipitation Mass Spectrometry Workflow for Large-scale Nuclear Interactome Profiling
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用于大规模核相互作用分析的无标签免疫沉淀质谱工作流程

15,307 Views

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11:19 min

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November 17, 2019

DOI:

10.3791/60432-v

DOI:
10.3791/60432-v

•

11:19 min
November 17, 2019

•

11 Views
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1Department of Molecular, Cellular and Developmental Biology,University of Colorado, 2Linda Crnic Institute for Down Syndrome,University of Colorado School of Medicine

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描述是一种蛋白质组学工作流程，用于利用感兴趣的特定蛋白质和无标签质谱的免疫亲和力来识别来自核亚细胞部分的蛋白质相互作用伙伴。该工作流程包括亚细胞分馏、免疫沉淀、过滤器辅助样品制备、离线清理、质谱和下游生物信息学管道。

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Label-freeImmunoprecipitationMass SpectrometryNuclear InteractomeProtein-protein InteractionsSubcellular FractionationAffinity PurificationProteomics WorkflowCell LysisNuclear LysateDounce Homogenization

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Guard, S. E., Ebmeier, C. C., Old, W. M. Label-Free Immunoprecipitation Mass Spectrometry Workflow for Large-scale Nuclear Interactome Profiling. J. Vis. Exp. (153), e60432, doi:10.3791/60432 (2019).

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