Assay Report Card

ID: ALA3431456

Type: Toxicity

Description: GSK_TCAKS: pIC50 HepG2 human cell line viability assay using luminiscence. Actively growing HepG2 cells were removed from a T-175 TC flask using 5mL Eagle's MEM (containing 10% FBS, 1% NEAA, 1% penicillin/streptomycin) and dispersed in the medium by repeated pipetting. Seeding density was checked to ensure that new mono-layers were not more than 50% confluent at the time of harvesting. Cell suspension was added to 500 mL of the same medium at a final density of 1.2 * 10^5 cells/mL. This cell suspension was dispensed (25 uL, 3000 cells per well) into 384-well clear-bottom plates using a Multidrop Combi dispenser. Prior to addition of the cell suspension, the screening compounds (250 nL) were pre-dispensed into the plates with an Echo liquid handler. Plates were incubated for 48 h at 37C, 5% CO2. After incubation, plates equilibrated at room temperature for 30 min before proceeding to develop the luminescent signal. The signal developer, CellTiter-Glo Reagent, was allowed to equilibrate at room temperature for 30 min and added to the plates (25uL per well) using a Multidrop Combi dispenser. Plates were left for 10 min at room temperature for stabilization and then read using a ViewLux.

Organism: Homo sapiens

Cell type: HepG2