Histone H2A.X Colorimetric Cell-Based ELISA Kit

A cell-based ELISA kit from Assay Genie offers a lysate-free, high-throughput format for qualitative detection of Histone H2A.X in cultured cells. The assay uses an indirect ELISA approach in which target-specific primary antibodies capture Histone H2A.X, and HRP-conjugated secondary antibodies bound to the Fc region of the primary antibody catalyse a colorimetric reaction upon substrate addition.

Detection is read at 450 nm, with a stated dynamic range of > 5000 cells and reactivity against Human, Mouse and Rat samples. A monoclonal antibody specific for human GAPDH is included as an internal positive control to normalise target absorbance values, while Crystal Violet whole-cell staining can be applied to determine cell density and correct for plating differences. The kit is configured in a 96-well microplate format and is stable at 4°C for 6 months.

Histone H2A.X replaces conventional H2A in a subset of nucleosomes and is a key mediator of the DNA damage response, required for checkpoint-mediated cell cycle arrest following low-dose ionising radiation and for efficient repair of DNA double-strand breaks. It is phosphorylated on S139 by ATM and DNA-PK in response to double-strand breaks generated by exogenous genotoxic agents or stalled replication forks, and its dysregulation has been implicated in cancer and neurodegenerative disorders, supporting its use as a relevant target for research and drug development.