Thermo Fisher, PV4548, LanthaScreen™ TR-FRET PPAR gamma Coactivator Assay Kit, goat

LanthaScreen™ TR-FRET PPAR gamma Coactivator Assay Kit, goat This kit contains Goat Tb-Anti-GST antibody; other kit components are the same as kit A15126:The LanthaScreen™ TR-FRET PPAR gamma Coactivator Assay Kit provides a sensitive and robust method for high-throughput screening of potential PPAR gamma ligands as agonists or antagonists of ligand-dependent coactivator recruitment. The kit uses a terbium (Tb)-labeled anti-GST antibody, a fluorescein-labeled coactivator peptide, and a Peroxisome Proliferator Activated Receptor (PPAR) gamma ligand-binding domain (PPAR gamma-LBD) that is tagged with glutathione-S-transferase (GST), in a homogenous mix-and-read assay format.
Agonist modeTo run the LanthaScreen™ TR-FRET Peroxisome Proliferator Activated gamma Receptor Coactivator Assay in agonist mode (to identify agonist compounds), PPAR gamma-LBD is added to ligand test compounds followed by addition of a mixture of the fluorescein-coactivator peptide and Tb-anti-GST antibody. After an incubation period at room temperature, the TR-FRET 520:495 emission ratio is calculated and used to determine the EC50 from a dose response curve of the compound. Based on the biology of the PPAR gamma-coactivator peptide interaction, this ligand EC50 is a composite value representing the amount of ligand required to bind to receptor, effect a conformational change, and recruit coactivator peptide (Figure 1).
Antagonist modeWhen the LanthaScreen™ TR-FRET Peroxisome Proliferator Activated gamma Receptor Coactivator Assay is run in antagonist mode (to identify antagonist compounds), PPAR gamma-LBD is added to ligand test compounds followed by addition of a mixture of agonist, fluores-cein-coactivator peptide, and Tb-anti-GST antibody. The concentration of agonist used in this mode is the EC80 concentration as determined by first running the assay in agonist mode (Figure 2).
Contents and Storage:The LanthaScreen™ TR-FRET PPARγ Coactivator Assay Kit contains PPAR gamma-LBD (GST) protein, fluorescently labeled TRAP220/ DRIP-2 coactivator peptide, Tb-anti-GST antibody, and buffers. Store components as indicated in the assay protocol (-80°C, -20°C, or +4°C).
Assay Entry: Biochemical coregulator interaction
Detection Method: Fluorescence
For Use With (Application): Co-Factor Interaction Assay, TR-FRET
For Use With (Equipment): Microplate Reader
Gene ID (Entrez): 5468
Ligand: PPAR Gamma
No. of Assays: 800 x 20 μL assays
Packaging: 384-well Plate
Product Type: TR-FRET PPAR Gamma Coactivator Assay Kit
Quantity: 800 x 20 μL assays
Readout: End Point
Shipping Condition: Dry Ice
Target Entry: PPARG, PPAR gamma, NR1C3
Conjugate: Tb (Terbium)
Product Line: LanthaScreen™
Unit Size: Each