Cells were plated in 96-well plate at the density of 1 × 104 per well and were allowed to attach for 24 h 5 μM Sorafenib (Cell Signaling Technology, USA), 5 μM ferrostatin-1 (S7243, Selleck, USA), 50 μM deferoxamine (S5742, Selleck, USA), 10 μM Z-VAD(OMe)-FMK (HY-16658, MedChemExpress, USA) or 1 μM Necrosulfonamide (HY-100573, MedChemExpress, USA)were added to the cells as indicated in Figure legends and incubated for 24 h. The cells were harvested and Cytotoxicity LDH assays were performed using the Cytotoxicity LDH assay kit (CK12, Dojindo, Japan) according the manufacturer's instructions as previously described [45 (link)].
Cytotoxicity ldh assay kit ck12
Manufactured by Dojindo Laboratories
Sourced in Japan
The Cytotoxicity LDH Assay Kit (CK12) is a product designed to measure lactate dehydrogenase (LDH) activity released from damaged cells. LDH is a stable cytoplasmic enzyme that is released upon cell lysis or damage, and its activity can be quantified colorimetrically to assess cytotoxicity.
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2 protocols using cytotoxicity ldh assay kit ck12
1
Cytotoxicity Evaluation of Compounds
2
Cytotoxicity and Apoptosis Assays
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Cells were seeded at 1 9 10 4 (for CAOV3 and ES2) or 6 9 10 3 (for JHOC5) cells/well in 96-well plates. The next day, medium was replaced and cells were treated with 10 nM FK866 for 36 h or left untreated. Lactate dehydrogenase (LDH) release was measured using a Cytotoxicity LDH Assay Kit (CK12) from Dojindo (Mashiki, Japan). Caspase 3/7 activity was measured using a Caspase-Glo 3/7 Assay Kit (G8090) from Promega (Madison, WI, USA). Results were normalized to cell numbers, as revealed by sulforhodamine staining of the replica plate.
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