Ldh cytotoxicity assay kit

Cytotoxicity was determined using the LDH-Cytotoxicity Assay Kit per manufacturer’s protocol (Abcam, Cambridge, MA).

SKOV3 cells were treated with rGO-Ag (0.2 µM) or TSA (0.2 µM) or combination of rGO-Ag (0.2 µM) and TSA (0.2 µM) for 24 h. The lactate dehydrogenase (LDH) leakage assay was carried out using the LDH cytotoxicity assay kit (Abcam). LDH activity was determined in the medium by measuring the absorbance at 490 nm using a Multiskan FC multiplate reader. A dead-cell protease activity assay was performed according to the method described by Jo et al.76 (link) CytoTox-Glo cytotoxicity assay (Promega Corp., Fitchburg, WI, USA) was performed to examine dead-cell protease activity by measuring the rGO-Ag-induced cytotoxicity in SKOV3. The cytotoxicity assay was employed to evaluate the cytotoxic effects of rGO-Ag (0.2 µM) or TSA (0.2 µM) or combination of rGO-Ag (0.2 µM) and TSA (0.2 µM). The cytotoxicity was determined by association of intracellular protease with a luminogenic peptide substrate (alanyl-alanylphenylalanyl-aminoluciferin). The degree of protease reaction can measure dead-cell protease activity. As a control, we treated cells with 1% Triton X-100 to exclude the background value of the medium color. Luminogenic peptide substrate (5 µL) was added to each well and incubated for 15 min at 37°C, and luminescence was measured using a luminescence counter (PerkinElmer) to determine the number of dead cells.