BDS CALUX - Bioassays
BDS offers an extensive panel of high-throughput-compatible CALUX reporter assays that enable effect identification and effect-based compound detection. Our assays allow for the sensitive and precise detection of specific biological effects at the cellular level. The panel contains assays relating to various effect classes such as endocrine disruption, xenobiotics metabolism, genotoxicity and acute toxicity. The influence of metabolic conversion of the samples may optionally be included in the test by the addition of metabolic enzymes. An overview of the CALUX assays that are currently available is presented here. CALUX analyses are performed in the BDS service laboratory, but are also run under license by others.
Assay Principle
Mammalian cells possess a wide range of mechanisms with which they can specifically respond to signals or stresses evoked by compounds they are confronted with. CALUX® (Chemically Activated LUciferase eXpression) assays form a panel of mammalian cell lines that were modified to produce a quantifiable response in addition to this natural response: If a cell is triggered to generate a response, a reporter gene (luciferase) is transcribed and translated into an enzyme that produces light during a reaction it catalyses. After addition of a substrate (luciferine), light production is quantified using a luminometer. The produced signal is proportional to the evoked effect. An overview of the CALUX® assays currently available is presented here.
Most cellular responses are mediated by a regulatory protein, which is a sensor that detects a certain event and subsequently incites the cellular response to this event. The regulator binds to a specific regulatory element which is present in DNA regions upstream of genes involved in the response. In the CALUX cell lines these regulatory elements were also placed in front of the luciferase (reporter) gene. Thereby, the production of the corresponding luciferase enzyme is coupled to the natural response of the cell.
Whereas most genes may be involved in multiple responses, the molecular design of the CALUX assays ensures that the observed response is highly specific for the effect of interest.
Assay Procedure
CALUX assays may be used for testing of compounds as well as complex mixtures. The cells are cultured in well plates (usually 96 or 384 wells), which allows for increased throughput and automated handling. Subsequently, they are exposed to a dilution series of the test compound or a dilution series of an extract of the test sample. Along with the test samples, the cells are also exposed to a concentration series of a reference compound. After a certain exposure time, the light production in the independent wells is quantified. The activity evoked by the test compound or test sample is subsequently derived by interpolation in the response curve of the reference compound and is expressed in equivalences of this reference compound.
Customer reviews
No reviews were found for BDS CALUX - Bioassays. Be the first to review!