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Adaptation of Hybridization Capture of Chromatin-associated Proteins for Proteomics to Mammalian Cells
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Genetics
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JoVE Journal Genetics
Adaptation of Hybridization Capture of Chromatin-associated Proteins for Proteomics to Mammalian Cells

Adaptation of Hybridization Capture of Chromatin-associated Proteins for Proteomics to Mammalian Cells

DOI:
10.3791/57140-v

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09:27 min

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June 01, 2018

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, , , , , , ,
1Department of Genetics,Texas Biomedical Research Institute, 2Department of Internal Medicine-Molecular Medicine,Wake Forest University School of Medicine, 3Department of Chemistry,University of Wisconsin

Chapters

  • 00:04Title
  • 00:44Cell Culture
  • 03:13Lysis and Shearing
  • 04:14Hybridization Capture
  • 06:14Evaluation of Capture Yield by qPCR with Primers for the Target Capture Region of Interest
  • 07:12Results: Hybridization Capture of Chromatin-associated Proteins in Mammalian Cells
  • 08:38Conclusion

Summary

Automatic Translation

Automatic Translation

This is a method to identify novel DNA-interacting proteins at specific target loci, relying on sequence-specific capture of crosslinked chromatin for subsequent proteomic analyses. No prior knowledge about potential binding proteins, nor cell modifications are required. Initially developed for yeast, the technology has now been adapted for mammalian cells.

Tags

Keywords: Hybridization CaptureChromatin-associated ProteinsProteomicsMammalian CellsGenome RegulationDNA Binding ProteinsCell CultureCross-linkingFormaldehydeGlycine

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