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FL-AR N-TERM Peptide

LanthaScreen® fluorescent nuclear receptor coregulator peptides contain known interaction motifs and are labeled with fluorescein. These peptides are matched and validated to complement the LanthaScreen® TR-FRET Nuclear Receptor Coregulator Assays. Assays developed using these reagents enable primary or secondary screening of nuclear receptor agonists and/or antagonists.

How they work
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. In the LanthaScreen® TR-FRET RXR beta Coactivator Assay, a terbium (Tb)-labeled anti-GST antibody is used to indirectly label GSTtagged RXR beta protein. An agonist (9-cis Retinoic Acid) added to the receptor which, upon ligand binding, causes a conformational change resulting in recruitment of the Fl-D22 coactivator peptide (Figure 1). The binding of the fluorescent coregulator peptide to RXR beta causes an increase in the TR-FRET emission ratio (Figure 2).

Contents and Storage:

LanthaScreen® fluorescent nuclear receptor coregulator peptides are supplied as 100 µM solutions in peptide-appropriate buffers. Store peptides at -20°C.

GeneBLAzer™ AR-UAS-bla Griptite™ Cells

AR-UAS-bla GripTite™ 293 cells contain the ligand-binding domain (LBD) of the rat Androgen receptor (AR) fused to the DNA-binding domain of GAL4 stably integrated in the GeneBLAzer®UAS-bla GripTite™ 293 cell line. GeneBLAzer®UAS-bla GripTite™ cells stably express a beta-lactamase reporter gene under the transcriptional control of an upstream activator sequence (UAS). When an agonist binds to the LBD of the GAL4 (DBD)-AR (LBD) fusion protein, the protein binds to the UAS, resulting in expression of beta-lactamase. Division Arrested (DA) cells are available in two configurations- an Assay Kit (which includes cells and sufficient substrate to analyze 1 x 384-well plate), and a tube of cells sufficient to analyze 10 x 384-well plates. AR-UAS-bla GripTite™ 293 cells are functionally validated for Z' and EC50 concentrations of R1881. Cells have been tested for assay performance under variable conditions, including DMSO concentration, cell number, stimulation time, and substrate loading time (data available upon request). Additional testing data using alternate stimuli are also available.

Androgen Receptor Monoclonal Antibody (AR 441) (Invitrogen™)

Androgen Receptor Monoclonal Antibody for Western Blot, IF, ICC, IHC (P), IP

Androgen Receptor Monoclonal Antibody (AR 441), Biotin (Invitrogen™)

Androgen Receptor Monoclonal Antibody for Western Blot, IF, ICC, IHC (P), IP

AR Monoclonal Antibody (OTI2E11), TrueMAB™ (OriGene)

AR Monoclonal Antibody for Western blot, IHC (P)

AR Monoclonal Antibody (OTI1D3), TrueMAB™ (OriGene)

AR Monoclonal Antibody for Western blot, IHC (P)

AR Monoclonal Antibody (UMAB240), UltraMAB™ (OriGene)

AR Monoclonal Antibody for IHC (P)

AR Monoclonal Antibody (OTI2H2), TrueMAB™ (OriGene)

AR Monoclonal Antibody for Western blot

AR Monoclonal Antibody (OTI3C5), TrueMAB™ (OriGene)

AR Monoclonal Antibody for Western blot, IHC (P)

AR Monoclonal Antibody (OTI2E6), TrueMAB™ (OriGene)

AR Monoclonal Antibody for Western blot, IHC (P)

Hamster IgG Isotype Control (Invitrogen™)

Hamster IgG Isotype Control for Flow, Ctrl

Androgen Receptor Recombinant Rat Protein, Ligand Binding Domain

Androgen receptor ligand binding domain (AR-LBD) is a 64.42 kDa recombinant rat protein, expressed as a GST- and His-tagged protein in insect cells. The protein contains the hinge region and an LBD amino acid sequence identical to that of the human LBD. Differences in the rat and human hinge region are present at human amino acids 646, 649, 650, 655, 656, 657, and 660. AR-LBD is expressed and purified in the absence of exogenous ligands, making it ideal for ligand-binding and coregulator recruitment studies.

AR-LBD rat recombinant protein is:

• Flexible—use in your in-house assay or as part of our PolarScreen™ Competitor assays for a complete assay solution
• Ready to use—active and tagged (with GST/His)
• Convenient—available in bulk sizes

Contact us for bulk quantities or for assistance with other nuclear receptor assay needs, from assay development to library screening and beyond.

LanthaScreen™ TR-FRET Androgen Receptor Coactivator Assay Kit

The LanthaScreen® TR-FRET Androgen Receptor Coactivator Assay Kit provides a sensitive and robust method for high-throughput screening of potential androgen receptor (AR) ligands as agonists of coactivator recruitment or antagonists of agonist-dependent coactivator recruitment. In both the agonist and antagonist modes, the homogeneous mix-and-read assay uses a rat AR ligand-binding domain (AR-LBD) tagged with hexahistidine and glutathione-S-transferase (GST) (also available separately), a terbium (Tb)-labeled anti-GST antibody, and a fluorescein-labeled coactivator peptide.

Agonist Mode
When the LanthaScreen TR-FRET Androgen Receptor Coactivator Assay is run in agonist mode (to identify agonist compounds), AR-LBD is added to ligand test compounds, followed by the addition of a mixture of fluorescein-coactivator peptide and Tb-anti-GST antibody. After room temperature incubation, the TR-FRET 520:495 nm emission ratio is calculated and used to determine the EC50 from a dose response curve of the compound. 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).

Antagonist Mode
When the LanthaScreen TR-FRET Androgen Receptor Coactivator Assay is run in antagonist mode (to identify antagonist compounds), AR-LBD is added to ligand test compounds, followed by addition of a mixture of DHT agonist, fluorescein-coactivator peptide, and Tb-anti-GST antibody. The concentration of DHT agonist used in this mode is the EC80 concentration as determined by first running the assay in agonist mode. An example of data produced in the antagonist mode is shown in the figure below.

beta-1 Adrenergic Receptor Polyclonal Antibody (Invitrogen™)

beta-1 Adrenergic Receptor Polyclonal Antibody for Western Blot, IF, ICC

Orion™ Star A Series Portable Meter Conductivity and DO Probe Holder (Thermo Scientific™)

Use the Thermo Scientific™ Orion™ Star A Series Portable Meter Conductivity and DO Probes Holder with the Star A series portable meter armor to protect your sensors in the field