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Label-Free Immunoprecipitation Mass Spectrometry Workflow for Large-scale Nuclear Interactome Profiling
JoVE 杂志
生物化学
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JoVE 杂志 生物化学
Label-Free Immunoprecipitation Mass Spectrometry Workflow for Large-scale Nuclear Interactome Profiling

Label-Free Immunoprecipitation Mass Spectrometry Workflow for Large-scale Nuclear Interactome Profiling

15,334 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

Summary

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Described is a proteomics workflow for identifying protein interaction partners from a nuclear subcellular fraction using immunoaffinity enrichment of a given protein of interest and label-free mass spectrometry. The workflow includes subcellular fractionation, immunoprecipitation, filter aided sample preparation, offline cleanup, mass spectrometry, and a downstream bioinformatics pipeline.

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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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