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For immunostaining analysis of apoptotic GAD67-positive cells, the In Situ Cell Death Detection Kit, Fluorescein (Roche, #11684795910) was used along with a polyclonal rabbit antibody to GAD67 (Abcam, #ab97739, 1:100) and anti-rabbit IgG Alexa Fluor 555 conjugated antibody (Invitrogen, # A31572, 1:400)

Usui I, Imamura T, Babendure JL, Satoh H, Lu JC, Hupfeld CJ, et al

Due to the evidently good predictivity of the in vivo alkaline comet assay, it seemed worthwhile to also evaluate the in vitro alkaline comet assay regarding its potential to detect mutagenic/carcinogenic NAs, when performed with relevant, metabolically competent mammalian tissue or cell types

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the membrane potential was held at -70 mV at room temperature (25C) during a period of 0.5-2 h after removal of the culture from the incubator