We offer a number of cell-based tissue culture assays, such as cell proliferation and cell cytotoxicity assays, flow cytometry assays (cell cycle, cell viability, membrane protein expression, TUNEL, caspase 3/7 apoptosis, etc). Standard cytotoxicity assays are based on the cleavage of the yellow terazolium salt (XTT) by metabolically active human cells (usually HeLa or A549 cells) to form an orange formazan dye. This conversion only occurs in viable cells, so cytotoxicity leads to a loss of activity. The formazan dye formed is soluble in aqueous solutions and is directly quantified using a scanning multi-well plate reader (spectrophotometer). Using multiple replicates, ensures that this method provides a high degree of accuracy and statistical data confidence. The assay is suitable for the performance of dose-response studies and is offered in a 96-well microplate format.
IC-50 for Tumor Cell Lines
IC-50 testing service is offered in over 120 in-house cell lines and primary cell types or any custom (client provided) cell line. The goal of this test is to measure the biological activity of a compound of interest (IC-50) against different types of tumor cell lines. In standard setup, cells in logarithmic growth are seeded in 96-well tissue culture plates (5,000-7,500 cells/well). It is important to use appropriate negative and positive controls in the assay (control conditions usually include the nontreated cells, cells with medium and vehicle as a negative control and one of the standard chemotherapy drugs as a positive control). Plates are assayed at 48-72 hr after initiation of compound exposure, then a 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) solution is added to each well for a 1- to 3-h incubation (at 37°C and 5% CO2). Analysis is performed at least in triplicate (biological replicate) and using a plate reader to generate IC-50 assay data.
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