@article{4ab1975ea54842c688086da785cb9f60,
title = "Detection and quantification of γ-H2AX using a dissociation enhanced lanthanide fluorescence immunoassay",
abstract = "Phosphorylation of the histone protein H2AX to form γ-H2AX foci directly represents DNA double-strand break formation. Traditional γ-H2AX detection involves counting individual foci within individual nuclei. The novelty of this work is the application of a time-resolved fluorescence assay using dissociation-enhanced lanthanide fluorescence immunoassay for quantitative measurements of γ-H2AX. For comparison, standard fluorescence detection was employed and analyzed either by bulk fluorescent measurements or by direct foci counting using BioTek Spot Count algorithm and Gen 5 software. Etoposide induced DNA damage in A549 carcinoma cells was compared across all test platforms. Time resolved fluorescence detection of europium as a chelated complex enabled quantitative measurement of γ-H2AX foci with nanomolar resolution. Comparative bulk fluorescent signals achieved only micromolar sensitivity. Lanthanide based immunodetection of γ-H2AX offers superior detection and a user-friendly workflow. These approaches have the potential to improve screening of compounds that either enhance DNA damage or protect against its deleterious effects.",
author = "Noubissi, {Felicite K.} and McBride, {Amber A.} and Leppert, {Hannah G.} and Millet, {Larry J.} and Xiaofei Wang and Davern, {Sandra M.}",
year = "2021",
month = apr,
day = "26",
doi = "10.1038/s41598-021-88296-3",
language = "English",
volume = "11",
pages = "8945",
journal = "Scientific Reports",
issn = "2045-2322",
publisher = "Nature Research",
number = "1",
}
