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Combined Immunofluorescence and DNA FISH on 3D-preserved Interphase Nuclei to Study Changes in 3D Nuclear Organization
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Biologia
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JoVE Journal Biologia
Combined Immunofluorescence and DNA FISH on 3D-preserved Interphase Nuclei to Study Changes in 3D Nuclear Organization

Combined Immunofluorescence and DNA FISH on 3D-preserved Interphase Nuclei to Study Changes in 3D Nuclear Organization

DOI:
10.3791/50087-v

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13:55 min

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February 03, 2013

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, ,
1Department of Pathology,New York University School of Medicine, 2New York University Center for Health Informatics and Bioinformatics, 3NYU Cancer Institute, 4Department of Pathology and Yale Cancer Center,Yale University School of Medicine

Capítulos

  • 00:05Título
  • 01:27DNA Probe Labeling with Fluorophores: Nick Translation
  • 03:37Probe Precipitation/Denaturation/Pre-annealing
  • 05:133-dimensional DNA FISH
  • 09:253-dimensional DNA FISH / Immuno-FISH
  • 10:40Combined Immunofluorescence and DNA FISH: Representative Results
  • 12:56Conclusion

Summary

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Here we describe a protocol for simultaneous detection of histone modifications by immunofluorescence and DNA sequences by DNA FISH followed by 3D microscopy and analyses (3D immuno-DNA FISH).

Tags

3D DNA FISH3D Nuclear Organization3D Confocal MicroscopyImmunofluorescenceChromatin ModificationsGene LociChromosome TerritoriesNuclear ArchitectureNuclear ProteinsHistone VariantsTranscription MachineryNuclear Sub-compartments

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