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Invitrogen™ LanthaScreen™ TR-FRET PPAR gamma Coactivator Assay Kit, goat

Catalog No. PV4548
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Quantity:
800 x 20 μL assays

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 mode
To 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 mode
When 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).
TRUSTED_SUSTAINABILITY

Specifications

Conjugate Tb (Terbium)
Detection Method Fluorescence
For Use With (Equipment) Microplate Reader
Quantity 800 x 20 μL assays
Packaging 384-well Plate
Content And Storage Store in ultra-cold freezer (-68 to -85°C).
Assay Entry Biochemical coregulator interaction
Gene ID (Entrez) 5468
Readout End Point
Shipping Condition Dry Ice
Target Entry PPARG, PPAR gamma, NR1C3
Product Line LanthaScreen™
For Use With (Application) Co-Factor Interaction Assay, TR-FRET
Ligand PPAR Gamma
Product Type TR-FRET PPAR Gamma Coactivator Assay Kit
No. of Assays 800 x 20 μL assays
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WARNING: Cancer - www.P65Warnings.ca.gov

For Research Use Only. Not for use in diagnostic procedures.

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