Understand the potential genotoxicity of your compound in our γH2AX double strand DNA damage response assay.
Our γH2AX double strand DNA damage response assay is in our portfolio of genotoxicity services. Cyprotex deliver consistent, high quality data and can adapt protocols based on specific customer requirements.
Taking into account the ability of the automated γH2AX assay to predict genotoxicity in vivo to the same accuracy as currently used in vitro assays, while its use of human metabolic competent cells, and its automated scoring, its limited use of test compound since small volumes are needed and its simple and rapid applicability to study large numbers of chemicals as it is amenable to robotised procedures, there are many arguments in favor of its usefulness as in vitro genotoxicity test. Especially its high sensitivity to detect DNA-reactive GTX compounds is a positive asset.
3Tsamou et al. (2012) Mutagenesis 27(6); 645-652
|Cell Line||HepG2 (other cell types available on request)|
|Multiplexing||Combination with other mechanistic endpoints available on request
Compatible with the in vitro HCS Micronucleus Test (MNT)
|Analysis Platform||Cellomics ArrayScan® VTI or XTI (Thermo Scientific)|
|Test Article Concentrations||8 point dose response curve with highest concentration based on cell loss or solubility limit (3 replicates per concentration)|
|Test Article Requirements||50 µL solution at 200x highest concentration or equivalent amount in solid|
|Time Points||In absence or presence of aroclor 1254 induced rat liver S9: 24 hr exposure time|
|Quality Controls||Negative control: 0.5% DMSO (vehicle)
Positive controls: Cyclophosphamide (S9 positive control) and chlorambucil (positive control)
|Data Delivery||Minimum effective concentration (MEC) and AC50 value for each measured parameter (cell loss, nuclear
morphology, DNA fragmentation and DNA damage)
|Minus Rat Liver S9||Plus Rat Liver S9|
|In vivo genotoxin|
|In vivo non-genotoxin|
1Mah LJ et al., (2010) γH2A.X: a sensitive molecular marker of DNA damage and repair. Leukemia 24; 679 – 686
2Chapman JR et al., (2012) Playing the End Game: DNA Double-Strand Break Repair Pathway Choice. Molecular Cell 47(4); 497-10
3Tsamou M et al., (2012) Performance of in vitro γH2A.X assay in HepG2 cells to predict in vivo genotoxicity. Mutagenesis 27(6); 645-652
4Diaz D et al., (2007) Evaluation of an automated in vitro micronucleus assay in CHO-K1 cells. Mutation Research 630;1-13
5Hastwell PW et al., (2009) Analysis of 75 marketed pharmaceuticals using the GADD45a-GFP ‘GreenScreen HC’ genotoxicity assay. Mutagenesis, 24(5); 455-463
Learn more about toxicology in our popular Mechanisms of Drug-Induced Toxicity guide