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Generation og oprensning af human INO80 Chromatin Remodeling komplekser og Subcomplexes
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JoVE Journal Biology
Generation and Purification of Human INO80 Chromatin Remodeling Complexes and Subcomplexes

Generation og oprensning af human INO80 Chromatin Remodeling komplekser og Subcomplexes

DOI:
10.3791/51720-v

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08:44 min

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October 23, 2014

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, , ,
1Stowers Institute for Medical Research, 2Department of Biochemistry & Molecular Biology,Kansas University Medical Center

Chapters

  • 00:05Title
  • 01:23Isolation of Nuclei
  • 02:45Extracting Nuclei with Salt
  • 04:38Immunoaffinity Purification of the Human INO80 or INO80 Subcomplexes
  • 06:53Results: Analysis of the Composition of Immunopurified INO80 Complexes and Subcomplexes
  • 08:01Conclusion

Summary

Automatic Translation

Automatic Translation

Denne protokol beskriver en fremgangsmåde til generering og oprensning af vildtype og mutant-versioner af humane INO80 kromatin remodellering kompleks. Epitop taggede versioner af INO80 underenheder stabilt udtrykt i HEK293-celler, og komplette komplekser og komplekser mangler specifikke sæt af underenheder oprenses ved immunaffinitetskromatografi.

Tags

INO80 Chromatin Remodeling ComplexNucleosome DynamicsDNA AccessibilityATP-dependent Nucleosome RemodelingIno80 ATPaseFLAG-tagged Ino80HEK 293 CellsSiRNA DepletionFLAG ImmunoprecipitationSubcomplexesMass SpectrometryChromatin Remodeling Assays

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