Cytotoxicity assays were performed in the presence of the anti-LSR mAb (#16-6), the mouse IgG2a isotype control antibody, the LSR-ADC or the control-ADC. The cytotoxicity of these agents was assessed by using cells growing (at a cell density of 500 to 4,000 cells per well) in a normal growth medium in flat-bottom 96-well white polystyrene plates (Thermo Fischer Scientific). After 24 h, the cells were incubated with serial dilutions of the agents in triplicate wells for 144 h, at 37°C, in a humidified 5% CO2 atmosphere. Cell viability was determined with the CellTiter-Glo Luminescent Cell Viability Assay Kit (Promega, WI), and luminescence values were measured by using a microplate reader (SPARK). The obtained results are reported as IC50; the concentration of compound needed to yield a 50% reduction in viability, as compared to vehicle-treated cells (control: 100%).
Flat bottom 96 well white polystyrene plates
Manufactured by Thermo Fisher Scientific
Flat-bottom 96-well white polystyrene plates are used for various laboratory applications. They provide a standardized, high-quality surface for performing experiments and assays. The plates are made of durable polystyrene material and have a flat bottom design to ensure consistent results.
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2 protocols using flat bottom 96 well white polystyrene plates
1
Cytotoxicity Assay for Anti-LSR mAb
2
Cytotoxicity Assay for GPC1-ADC
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Cytotoxicity assays performed in the presence of GPC1-ADC, or control-ADC were assessed using cells plated at 2,500 (KKU-055 and KKU-055 GPC1 KO) or 1,000 (SSP-25 and TFK-1) cells per well in normal growth medium in flat bottom 96-well white polystyrene plates (Thermo Fisher Scientific). After 24 hours, tumor cells were added with serial dilutions of GPC1-ADC or control-ADC in triplicate wells and incubated for 144 hours at 37 C in a humidified 5% CO 2 atmosphere. Cell viability was determined using the CellTiter-Glo Luminescent Cell Viability Assay Kit (Promega), as described previously (26) .
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