The Alfred Medical Research and Education Precinct

4 ARTICLES PUBLISHED IN JoVE

Biology

Quantification of γH2AX Foci in Response to Ionising Radiation

Li-Jeen Mah^1,2, Raja S. Vasireddy^1,2,3, Michelle M. Tang^1,3, George T. Georgiadis¹, Assam El-Osta^2,3, Tom C. Karagiannis^1,2

¹Epigenomic Medicine, Baker IDI Heart and Diabetes Institute, The Alfred Medical Research and Education Precinct, ²Department of Pathology, The University of Melbourne, ³Epigenetics in Human Health and Disease, Baker IDI Heart and Diabetes Institute, The Alfred Medical Research and Education Precinct

Quantification of DNA double-strand streaks using γH2AX formation as a molecular marker has become an invaluable tool in radiation biology. Here we demonstrate the use of an immunofluorescence assay for quantification of γH2AX foci after exposure of cells to radiation.

Biology

Quantitation of γH2AX Foci in Tissue Samples

Michelle M. Tang^1,2, Li-Jeen Mah^1,3, Raja S. Vasireddy^1,2,3, George T. Georgiadis¹, Assam El-Osta^2,3, Simon G. Royce^3,4,5, Tom C. Karagiannis^1,3

¹Epigenomic Medicine, Baker IDI Heart and Diabetes Institute, The Alfred Medical Research and Education Precinct, ²Epigenetics in Human Health and Disease, Baker IDI Heart and Diabetes Institute, The Alfred Medical Research and Education Precinct, ³Department of Pathology, The University of Melbourne, ⁴Department of Allergy and Immunology, Murdoch Children's Research Institute, Royal Children's Hospital, ⁵Department of Pediatrics, The University of Melbourne

Quantitation of DNA double-strand breaks on the basis of γH2AX foci has become an invaluable tool, particularly in radiation biology, for the evaluation of tissue radiosensitivity and effects of radiation modifying compounds. Here we demonstrate the use of an immunofluorescence assay for quantitation of γH2AX foci in tissue samples.

Biology

Evaluation of the Spatial Distribution of γH2AX following Ionizing Radiation

Raja S. Vasireddy^1,2,3, Michelle M. Tang^1,2, Li-Jeen Mah^2,3, George T. Georgiadis², Assam El-Osta^1,3, Tom C. Karagiannis^2,3

¹Epigenetics in Human Health and Disease, BakerIDI Heart and Diabetes Institute, The Alfred Medical Research and Education Precinct, ²Epigenomic Medicine, BakerIDI Heart and Diabetes Institute, The Alfred Medical Research and Education Precinct, ³Department of Pathology, University of Melbourne

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.

Biology

Clonogenic Assay: Adherent Cells

Haloom Rafehi^1,2, Christian Orlowski^1,2,3, George T. Georgiadis¹, Katherine Ververis^1,4, Assam El-Osta^2,3, Tom C. Karagiannis^1,2

¹Epigenomic Medicine, BakerIDI Heart and Diabetes Institute, The Alfred Medical Research and Education Precinct, ²Department of Pathology, The University of Melbourne, ³Epigenetics in Human Health and Disease, BakerIDI Heart and Diabetes Institute, The Alfred Medical Research and Education Precinct, ⁴Department of Anatomy and Cellular Biology, The University of Melbourne

The applicability of the clonogenic assay for evaluating reproductive viability has been established for more than 50 years. Here we demonstrate the general procedure for performing the clonogenic assay with adherent cells.