Cells (20,000 per well) are grown for 24 h in 96-well plates, then exposed to 2-HBA (bis(2-hydroxybenzylidene)acetone) for either 24 h (for glutathione determination) or 48 h (for determination of enzyme activities). At the end of the exposure period, cells are collected by centrifugation (1500×g for 15 min at 4°C), washed with DPBS, and finally lysed in 0.08% digitonin. An aliquot (25 μL) is used for protein analysis. Activity of NQO1 is determined by the Prochaska test^[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

After exposure to 2-HBA (bis(2-hydroxybenzylidene)acetone) for 24 h, duplicate aliquots of cells (1×10⁶) are collected by centrifugation and washed with cold DPBS. Apoptosis is determined using the Annexin-V-FLUOS assay with simultaneous determination of the necrotic fraction by the uptake of propidium iodide^[1].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Dinkova-Kostova AT, et al. Bis(2-hydroxybenzylidene)acetone, a potent inducer of the phase 2 response, causes apoptosis in mouse leukemia cells through a p53-independent, caspase-mediated pathway. Cancer Lett. 2007 Jan 8;245(1-2):341-9.

266.29

C₁₇H₁₄O₃

131359-24-5

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