"TR-FRET" Related Products

TR-FRET CAR Coactivator Assay Kit, rabbit

Cat.No.: Kit-0892
Description: The TR-FRET Constitutive Androstane Receptor (CAR) Coactivator Assay provides a sensitive and robust method for highthroughput screening (HTS) of potential CAR ligands as agonists of ligand-dependent coactivator recruitment or inverse agonists of liganddependent coactivator displacement. The kit uses a terbium-labeled anti-GST antibody, a fluorescein-labeled coactivator peptide, and a CAR ligand-binding domain (CAR-LBD) that is tagged with glutathione-S-transferase (GST) in a homogeneous mix-and-read assay format.
To assay:
When running the TR-FRET CAR Coactivator Assay, CAR-LBD is added to ligand test compounds followed by addition of a mixture of the fluorescein-coactivator peptide and terbium anti-GST antibody. After an incubation period at room temperature, the 520 nm/ 495 nm TR-FRET ratio is calculated and can be used to determine the EC50 from a dose response curve of the compound. Based on the biology of the CAR-coactivator peptide interaction, this ligand’s EC50 is a composite value representing the amount of ligand required to bind to receptor, effect a conformational change, and either recruit or displace coactivator peptide.
Applications: Nuclear Receptor Assay
Storage: Tb-anti-GST Antibody (Rabbit): Store at -20°C
CAR-LBD, GST: Store at -80°C
Fluoroscein Coactivator Peptide: Store at -20°C
TR-FRET Coregulatore Buffer G: Store at 4°C
1M DTT: Store at -20 °C or -80 °C
Size: 800 x 20 µL assays
Materials Required but Not Supplied: Microplate Reader
Detection method: Fluorescent
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Optional requirements on this protein    +Expand
C-fusion    N-fusion   Non-tagged
His    GST   Fc   Others
<1.0 eu/μg    <0.1 eu/μg   <0.01 eu/μg   Not required
Monomer Isolation    Dimer Isolation    Not required
>80% by SDS-PAGE    >90% by SDS-PAGE   >95% by SDS-PAGE   Others

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