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Plasmids for CALUX Bioassays Chemical Activated LUciferase gene eXpression (CALUX) is a ligand-dependent nuclear receptor-based bioassay method developed to screen and relatively quantify chemicals or classes of chemicals, such as dioxins / dioxin-like compounds, in samples. CALUX bioassays have been widely used as tools by research and commercial laboratories for detection of diverse chemicals of interest. This method is based on an engineered stable cell line that containing a reporter gene (e.g., luciferase, SEAP, GFP) under the control of receptor-specific DNA binding sites. When a CALUX cell line is triggered to generate a response, a reporter gene is transcribed and translated which can be easily detected. The produced signal is proportional to the evoked effect.  A brief general assay procedure is as follows: 1. Thaw and culture a CALUX cell line at 37⁰C with 5% CO2 using appropriate culture medium, e.g., DMEM, 10% Fetal Bovine Serum (FBS).2. Harvest and seed cells into a 96-well microplate in 100 µl of growth medium with preferred cell number per well (e.g., 10000 cells/well). 3. Incubate cells at 37⁰C in a CO2 incubator for overnight. 4. The next day, stimulate cells with a dilution series of the test samples. Along with the test samples, the cells are also treated with a dilution series of a reference compound. 5. Incubate cells at 37⁰C in a CO2 incubator for a certain time (e.g., 16~72 hours). 6. Detect the reporter gene expression and plot the dose-responsive curve 7. Data interpretation RGBiotech offers a series of reporter plasmids that can be used for generating CALUX reporter cell lines. |
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