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

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

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).

Conjugate

Tb (Terbium)

Detection Method

Fluorescent

For Use With (Equipment)

Microplate Reader

Packaging

384-well Plate

Content And Storage

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

Assay Entry

Biochemical coregulator interaction

Gene ID (Entrez)

5465

Readout

End Point

Shipping Condition

Dry Ice

Target Entry

PPARA, PPAR alpha, NR1C1

Product Line

LanthaScreen™

For Use With (Application)

Co-Factor Interaction Assay, TR-FRET

Ligand

PPAR alpha

Product Type

TR-FRET PPAR Alpha Coactivator Assay Kit

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