规格
Assay Entry:	Biochemical coregulator interaction
Product Line:	LanthaScreen®
Readout:	End Point
Shipping Condition:	Dry Ice
Target Entry:	RXRA, RXR alpha, NR2B1
Technique:	TR-FRET
Validated Application:	Co-Factor Interaction Assay
Label or Dye:	Tb (Terbium)
Detection Method:	Fluorescent
For Use With (Equipment):	Microplate Reader
Format:	384-well plate

储存
LanthaScreen® TR-FRET RXR alpha Coactivator Assay Kits contain RXR 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).

描述

This kit contains Goat Tb-Anti-GST antibody; other kit components are the same as kit A15138:

The LanthaScreen® TR-FRET Retinoid X Receptor alpha Coactivator Assay provides a sensitive and robust method for high-throughput screening of potential RXR alpha ligands as agonists of ligand-dependent coactivator recruitment. The kit uses a terbium (Tb)-labeled anti- GST antibody, a fluorescein-labeled coactivator peptide, and a human recombinant RXR alpha ligand-binding domain (RXR alpha-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 Retinoid X Receptor alpha Coactivator Assay in agonist mode (to identify agonist compounds), RXR 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 TR-FRET ratio of 520/495 is calculated and can be used to determine the EC50 from a dose-response curve of the compound. Based on the biology of the RXR 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).

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