This kit contains Goat Tb-Anti-GST antibody; other kit components are the same as kit A15144:
The LanthaScreen™ TR-FRET PPAR delta Competitive Binding Assay provides a sensitive and robust method for high-throughput screening (HTS) of ligands for peroxisome proliferator-activated receptor delta (PPAR delta). The kit uses a terbium-labeled anti-GST antibody, a fluorescent small-molecule pan-PPAR ligand (Fluormone™ Pan-PPAR Green), and a human PPAR delta ligand-binding domain (LBD) that is tagged with glutathione S-transferase (GST), in a homogeneous mix-and-read assay format.
To assay: When running the LanthaScreen™ TR-FRET PPAR delta Competitive Binding Assay, Fluormone™ Pan-PPAR Green is added to ligand test compounds, followed by addition of a mixture of the PPAR delta-LBD and terbium anti-GST antibody. When the Fluormone™ Pan-PPAR Green is bound to PPAR delta, energy transfer from the terbium-labeled antibody to the tracer occurs, and a high TR-FRET ratio is observed. Competitive ligand binding to PPAR delta is detected by a test compound’s ability to displace the tracer, which results in a loss of FRET between the antibody and the tracer. After an incubation period at room temperature, the 520 nm/495 nm TR-FRET ratio is calculated and can be used to determine the IC50 from a dose response curve of the compound (Figure 1). This type of binding assay is analogous to radioligandbased assays, except that it eliminates handling of radioactivity and enables a homogeneous, "addition-only" format.
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