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

LanthaScreen® TR-FRET PPAR alpha Coactivator Assay Kit, goat

Catalog number: PV4684

 Related applications: Target & Lead Identification & Validation

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Description

This kit contains Goat Tb-Anti-GST antibody; other kit components are the same as kit A15133:

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

Specifications

Format: 384-well plate
Gene ID: 5465
Readout: End Point
Technique: TR-FRET
Label or Dye: Tb (Terbium)
Product Size: 800 x 20 µL assays
Detection Method: Fluorescent
Validated Application: Co-Factor Interaction Assay
For Use With (Equipment): Microplate Reader
Shipping Condition: Dry Ice

Contents & storage

Store in ultra-cold freezer (-68 to -85°C).

Documents

Manuals & protocols