Ligand binding to nuclear receptors causes conformational changes in the receptor, resulting in a cascade of events, including dissociation of repressor proteins,association of coactivator proteins, and assembly of pol II and other transcriptional factors for activation of target genes. TR-FRET based assays can be developed using the LanthaScreen™ panel of fluorescein-labeled coregulator peptides to investigate conformational changes of nuclear receptors upon ligand binding, either by determining the affinity of ligand- bound receptor for different coregulator peptides, or by identifying additional agonists or antagonists via displacement or recruitment of a specific coregulator peptide. 

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LanthaScreen™ TR-FRET NR Coregulator Interaction Assay Kits contains the reagents necessary to perform an assay to assess the ligand dose dependency for the recruitment of coregulator peptides to nuclear receptor targets.

LanthaScreen™ fluorescent nuclear receptor coregulator peptides contain known interaction motifs and are labeled with fluorescein. Assays using these reagents enable primary or secondary screening of nuclear receptor agonists and/or antagonists.


 

Figure 1. Principle of nuclear receptor agonist-dependent coactivator peptide recruitment. Tb-anti-GST antibody indirectly labels RXR-β by binding to the GST tag on the N-terminus. Binding of the agonist, 9-cis-RA, to RXR-β causes a conformational change that results in an increase in the affinity of RXR-β for fluorescein-D22, a labeled coactivator peptide. The close proximity of the fluorescently labeled coactivator peptide to the Tb-labeled antibody causes an increase in the TR-FRET signal.

 

 

Figure 2. Representative data measuring recruitment of fluorescein-D22 coactivator peptide to RXR-β. Serial dilution of agonist 9-cis retinoic acid, 1 nM RXR β- LBD, 350 nM fluorescein-D22 peptide, and 2 nM Tb-anti-GST antibody. Results for 1-hr, 2-hr, 4-hr, and 6-hr incubations are shown with the corresponding EC50 values. The curves were generated using a sigmoidal dose response (variable slope) equation in GraphPad™ Prism® 4.0.

 

Ordering Information

Sku Name Size Price Qty
PV4836 LanthaScreen® TR-FRET CAR Coactivator Assay Kit, goat 800 x 20 µL assays USD 1,358.00
PV4408 LanthaScreen® TR-FRET ERR gamma Coactivator Assay Kit, goat 800 x 20 µL assays USD 1,358.00
PV4409 LanthaScreen® TR-FRET RAR alpha Coactivator Assay Kit, goat 800 x 20 µL assays USD 1,358.00
PV4655 LanthaScreen® TR-FRET LXR alpha Coactivator Assay Kit, goat 800 x 20 µL assays USD 1,358.00
PV4684 LanthaScreen® TR-FRET PPAR alpha Coactivator Assay Kit, goat 800 x 20 µL assays USD 1,358.00
PV4411 LanthaScreen® TR-FRET RAR gamma Coactivator Assay Kit, goat 800 x 20 µL assays USD 1,358.00
PV4548 LanthaScreen® TR-FRET PPAR gamma Coactivator Assay Kit, goat 800 x 20 µL assays USD 1,358.00
PV4685 LanthaScreen® TR-FRET PPAR delta Coactivator Assay Kit, goat 800 x 20 µL assays USD 1,358.00
PV4658 LanthaScreen® TR-FRET LXR beta Coactivator Assay Kit, goat 800 x 20 µL assays USD 1,358.00
PV4686 LanthaScreen® TR-FRET TR beta Coactivator Assay Kit, goat 800 x 20 µL assays USD 1,358.00
PV4687 LanthaScreen® TR-FRET TR alpha Coactivator Assay Kit, goat 800 x 20 µL assays USD 1,358.00
PV4688 LanthaScreen® TR-FRET RAR beta Coactivator Assay Kit, goat 800 x 20 µL assays USD 1,358.00
PV4663 LanthaScreen® TR-FRET ERR alpha Coactivator Assay Kit, goat 800 x 20 µL assays USD 1,358.00
PV4797 LanthaScreen® TR-FRET RXR alpha Coactivator Assay Kit, goat 800 x 20 µL assays USD 1,358.00
PV4541 LanthaScreen® TR-FRET ER beta Coactivator Assay Kit, goat 800 x 20 µL assays USD 1,358.00
PV4800 LanthaScreen® TR-FRET ERR beta Coactivator Assay Kit, goat 800 x 20 µL assays USD 1,358.00
PV4544 LanthaScreen® TR-FRET ER alpha Coactivator Assay Kit, goat 800 x 20 µL assays USD 1,358.00
PV4385 LanthaScreen® TR-FRET RXR beta Coactivator Assay Kit, goat 800 x 20 µL assays USD 1,358.00
PV4666 LanthaScreen® TR-FRET PR Coactivator Assay Kit, goat 800 x 20 µL assays USD 1,358.00
PV4554 LanthaScreen® TR-FRET VDR Coactivator Assay Kit, goat 800 x 20 µL assays USD 1,358.00
PV4833 LanthaScreen® TR-FRET FXR Coactivator Assay Kit, goat 800 x 20 µL assays USD 1,358.00
PV4683 LanthaScreen® TR-FRET GR Coactivator Assay Kit, goat 800 x 20 µL assays USD 1,358.00
PV4381 LanthaScreen® TR-FRET AR Coactivator Assay Kit, goat 800 x 20 µL assays USD 1,358.00
PV5890 LanthaScreen® TR-FRET ROR gamma Coactivator Assay Kit, goat 800 x 20 µL assays USD 1,388.00
PV5889 LanthaScreen® TR-FRET ROR alpha Coactivator Assay Kit, goat 800 x 20 µL assays USD 1,388.00

For research use only. Not intended for any animal or human therapeutic or diagnostic use.