In mammalian cells, a double-strand break (DSB) in genomic DNA is potentially lethal to a cell, and one of the responses to DSB formation as part of the repair process is phosphorylation of histone H2AX.
See product information for DNA damage assays
DNA damage can be measured as an indicator of genotoxicity using an antibody against phosphorylated H2AX. By combining specific antibody-based detection of DNA damage with a cytotoxicity indicator, both parameters can be measured simultaneously in the same cell.
The HCS DNA Damage Kit uses a secondary antibody conjugate to detect phosphorylated H2AX, Image-iT™ DEAD Green™ dye to detect cytotoxicity, and Hoechst™ 33342 to label nuclei in both live and dead cells. The kit is compatible with additional multiplexing steps, and results can be analyzed using automated image analysis techniques.
Using the HCS DNA Damage Kit, researchers can detect and quantitate DNA damage and changes in cell permeability.
|Readout||Combined genotoxicity and cytotoxicity detection compatible with automated image analysis in HCS|
|Common filter set||DAPI||FITC||TRITC|
|Reporter||Hoechst 33342 stain||Image-iT DEAD Green dye||Alexa Fluor 555 dye|
|Added to growth media||No||Yes||No|
|Formaldehyde fixable||Cells are labeled after fixation||Yes||Cells are labeled after fixation|
|Multiplexing||Can be multiplexed with additional detection reagents|
|Format||1 kit (2 x 96-well plates)|
|Molecular Probes™ Handbook
||BioProbes™ journal articles