GADD45a mediates the adaptive response to genotoxic stress. The patented GLuc reporter includes complex regulatory elements, and gives positive results for direct acting mutagens, clastogens, as well as aneugens, and topoisomerase and polymerase inhibitors. Importantly the assay gives correct negative results for non-genotoxins, many of which give misleading positive results in other in vitro tests.
Without metabolic activation (–S9)
Eight, 2-fold dilutions of each compound together with positive controls are set out in the 96well microplate and growing cells are added to each well. After incubation for 48 hours and measurement of the flash luminescence in a microplate reader equipped with an auto-injection facility, simple software gives automated decisions and a clear graphical output.
With metabolic activation (+S9)
Cells are incubated in 96
A unique combination of both high specificity and high sensitivity genotoxicity assessment in a human cell line has been demonstrated in a wide range of validation exercises with diverse sets of compounds. See the literature section (below, right) for details of specific publications.
The BlueScreen Assay has recently been adapted for automation compatibility and hence use in 384-well format