Cells are grown in multi-well plates (A) and incubated with compound for 72-hours (B, C). The number of viable cells is determined with CellTiter-Glo based on the amount of ATP present (D).
A tyrosine kinase inhibitor and staurosporine were tested for proliferation inhibition of 12 cancer cell lines. 72 h after compound addition, live cells were quantified via CellTiter-Glo. IC50 values are expressed as a percentage of proliferation in the presence of solvent alone (100% = high control) compared with cells treated with 1E-5M staurosporine (0% = low control).
Combining drugs that affect synergistically-acting targets is a promising strategy for drug discovery.
Depending on the drugs’ effectiveness and their mechanisms of action, different approaches are being used to determine combinatorial drug effects.
At Reaction Biology, we can perform
(i) a one-sided approach, i.e., the concentration of one drug is set, and the concentration of the other drug changes or
(ii) a two-sided approach, i.e., both drugs are tested with several concentrations in a checkerboard matrix setup.
Combination testing is available for Cell Proliferation Assay, ProLiFiler, Soft Agar Assay, Migration Assay, and 3D Tumor Spheroid Assay.
Assay details for CellTiter-Glo readout.
Setup: IC50 value determination and single concentration testing. Free choice for screening of the cell lines of your choice or panel screening via ProLiFiler.
Controls: DMSO only treatment serve as high control. Staurosporine treatment (at 1x10-5M) serves as low control. In every project, testing of a standard reference compound is included in IC50 format.
Custom assay development: Custom-tailored assays can be performed. Cell lines that are not part of the list, can be established according to the client’s research needs.
Report: A detailed report including assay conditions, methodology, and comprehensive evaluation of data as well as raw data for each analysis will be provided.
Turnaround time for standard studies is about 3 weeks. Expedited scheduling and data delivery can be arranged prior to study commencement.
Screening facility: CellTiter-Glo assays are performed at our German site. Additional readout formats are performed at our US site.