This kit contains a rabbit Tb-anti-GST antibody—an equivalent kit containing a goat-sourced antibody is also available.

LanthaScreen® TR-FRET PPAR gamma Coactivator Assay Kit, rabbit

Catalog number: A15126

 Related applications: Pharma & Biopharma | Target & Lead Identification & Validation

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Description

This kit contains Rabbit Tb-Anti-GST antibody; other kit components are the same as kit PV4548:

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

Specifications

Color: Green
Detection Method: Fluorescent
Druggable Target: Nuclear Receptors
For Use With (Equipment): Microplate Reader
Format: 384-well plate
Label or Dye: Tb (Terbium)
Product Line: LanthaScreen®
Product Size: 800 x 20 µL assays
Readout: TR-FRET (Activity)
Technique: TR-FRET
Validated Application: Nuclear Receptor Assay

Contents & storage

Tb-anti-GST Antibody (Rabbit): Store at -20 °C
PPAR gamma-LBD, GST: Store at -80 °C
Fluoroscein Coactivator Peptide: Store at -20 °C
TR-FRET Coregulatore Buffer F: Store at 4 °C
1M DTT: Store at -20 °C or -80 °C

Documents

Manuals & protocols