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Chapters
- 00:00Title
- 00:05Introduction
- 01:01Cell Preparation
- 02:28Immunofluorescence Staining
- 05:00Microscopy
- 06:12Analysis Foci Counting
- 07:56Analysis Line Scan
- 08:39Analysis 3D-Image Reconstruction
- 09:09Conclusion
Microscopic analysis of γH2AX foci, which form following the phosphorylation of H2AX at Ser-139 in response to DNA double-strand breaks, has become an invaluable tool in radiation biology. Here we used an antibody to mono-methylated histone H3 at lysine 4 as an epigenetic marker of actively transcribing euchromatin, to evaluate the spatial distribution of radiation-induced γH2AX formation within the nucleus.
Tags
Spatial DistributionγH2AXIonizing RadiationDNA Double-strand Breaks (DSBs)Ser-139 ResidueHistone H2AXPhosphorylationPhosphatidyl-inosito 3-kinase (PI3K) FamilyAtaxia Telangiectasia Mutated (ATM)DNA-protein Kinase Catalytic SubunitATM And RAD3-related (ATR)Immunofluorescence MicroscopyγH2AX FociDSB Formation And RepairRadiation Modifying CompoundsCytotoxic CompoundsDNA Damage And RepairChromatin Templateγ-irradiation-induced γH2AX Foci
