Thermo Scientific™ LanthaScreen™ TR-FRET PPAR alpha Coactivator Assay Kit, rabbit

Product Code. 15343796

Brand: Thermo Scientific™ A15133

Description

The LanthaScreen™ TR-FRET PPAR (peroxisome proliferator activated receptor) alpha Coactivator Assay provides a sensitive and robust method for high-throughput screening of potential PPAR alpha ligands as agonists or antagonists of ligand-dependent coactivator recruitment.

This kit contains Rabbit Tb-Anti-GST antibody; other kit components are the same as kit PV4684:

The LanthaScreen™ TR-FRET PPAR (peroxisome proliferator activated receptor) alpha Coactivator Assay provides a sensitive and robust method for high-throughput screening of potential PPAR alpha 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 human recombinant PPAR alpha ligand-binding domain (PPAR alpha-LBD) that is tagged with glutathione-S-transferase (GST) in a homogeneous mix-and-read assay format.

Agonist mode:

When running the LanthaScreen™ TR-FRET PPAR alpha Coactivator Assay in agonist mode (to identify agonist compounds), PPAR alpha- LBD is added to ligand test compounds followed by addition of a mixture of the fluorescein-coactivator peptide and Tb-labeled 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 PPAR alpha-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 (see Figure 1).

Antagonist mode:

When the LanthaScreen™ TR-FRET PPAR alpha Coactivator Assay is run in antagonist mode (to identify antagonist compounds), PPAR alpha-LBD is added to ligand test compounds followed by addition of a mixture of agonist, fluorescein-coactivator peptide, and Tb-labeled anti-GST antibody (Figure 2). The concentration of agonist used in this mode is the EC80 concentration as determined by first running the assay in agonist mode.

Contents and Storage:

The LanthaScreen™ TR-FRET PPAR alpha Coactivator Assay Kit contains PPAR alpha-LBD (GST) protein, fluorescently labeled PGC1a coactivator peptide, Tb-labeled anti-GST antibody, and buffers. Store components as indicated in the assay protocol (-80°C, -20°C, or 4°C).

Specifications

• Color: Green
• Conjugate: Tb (Terbium)
• Description: LanthaScreen™ TR-FRET PPAR Alpha Coactivator Assay Kit, 800 x 20μL Assays, Green, 384-Well Plate Format, Tb (Terbium) Dye Type, Fluorescent Detection Method, TR-FRET Technique, For Pharma & Biopharma, Target Lead Identification & Validation
• Detection Method: Fluorescence
• For Use With (Equipment): Microplate Reader
• Includes: Tb-anti-GST antibody (rabbit), PPAR alpha-LBD, GST, fluorescein coactivator peptide, TR-FRET coregulator buffer J, 1M DTT
• Quantity: 800 x 20 μL assays
• Regulatory Status: For research use only (RUO)
• Packaging: 384-well Plate
• Content And Storage: • Tb-anti-GST Antibody (Rabbit) (store at -20°C)
  • PPAR alpha-LBD, GST (-80°C)
  • Fluoroscein Coactivator Peptide (-20°C)
  • TR-FRET Coregulatore Buffer J (4°C)
  • 1M DTT (-20°C or -80°C)
• Assay Category: Biochemical Assay
• Druggable Target: Nuclear Receptors
• Readout: TR-FRET (Activity)
• Product Line: LanthaScreen™
• For Use With (Application): Nuclear Receptor Assay, TR-FRET
• Ligand: PPAR alpha
• Product Type: TR-FRET PPAR Alpha Coactivator Assay Kit
• No. of Assays: 800 x 20 μL Assays

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