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

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

描述

This kit contains Rabbit Tb-Anti-GST antibody; other kit components are the same as kit PV4833:

The LanthaScreen® TR-FRET FXR (farnesoid X receptor) Coactivator Assay Kit provides a sensitive and robust method for high-throughput screening of potential FXR ligands as agonists of coactivator recruitment. The kit uses a terbium (Tb)-labeled anti-GST antibody, a fluorescein- labeled coactivator peptide, and an FXR ligand-binding domain (FXR-LBD) that is tagged with glutathione-S-transferase (GST), in a homogeneous, mix-and-read assay format.

To assay:
When running the LanthaScreen® TR-FRET FXR coactivator assay in agonist mode, FXR-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 used to determine the EC50 from a dose response curve of the compound. Based on the biology of the FXR-coactivator peptide interaction, the 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).

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