Ab65393 ldh cytotoxicity assay kit

Cytotoxicity was assessed based on the release of lactate dehydrogenase (LDH) using a cytotoxicity detection kit (ab65393 LDH Cytotoxicity Assay Kit, Abcam, USA), a colorimetric assay for the quantification of cell death based on the measurement of lactate dehydrogenase activity released from the cytosol of damaged cells into the supernatant. Primary neurons were seeded at a density of 6×10⁴ cells per well in 96 well plate for 7 days. After experimental treatment, each well was mixed with LDH reaction buffer and gentle shaking at room temperature for 30 min. Cells without any treatment served as low controls. A high control was treated with the cell lysis solution. Each sample was assayed in triplicate, and the wavelength was measured at 450 nm. To determine the percentage of cytotoxicity, the average absorbance values of the triplicate readings were calculated, and the values were substituted into the following equation: Cytotoxicity (%) = (test sample value – low control value / high control value – low control value) × 100.

Cell cytotoxicity was determined by measuring the lactate dehydrogenase (LDH) release (ab65393, LDH-Cytotoxicity Assay kit, Abcam) from iPSC ioNEURONS/glut under different treatment conditions. Ten μl supernatant samples from the neuronal cultures under different conditions were collected and processed following the manufacturer’s protocol. The LDH released was determined by measuring the absorbance of the samples with a Synergy H1 microplate reader (BioTek) at 450 nm.