Every PolarScreen™ Competitive Binding Assay for Nuclear Receptors includes protein, a proprietary fluorescent Fluormone™ ligand, and an optimized buffer system. When the NR binds to the Fluormone™ ligand, the resulting complex yields a high polarization value. If the test compound displaces the Fluormone™ ligand from the complex, the polarization value is lowered. Since this occurs only in the presence of a test compound, the shift in polarization value enables you to accurately and conveniently determine relative affinity of a test compound for the NR.

PolarScreen™ Competitive Binding Assays for Nuclear Receptors

   
Figure 1. Fluormone™ ligands are biologically relevant steroid derivatives, designed to achieve nanomolar binding affinities and robust Z’-factors. Development of these Fluormone™ ligands required expert chemistry and manipulation of buffer composition. Assay are optimized for maximum stability of instrumentation platforms, and to tolerate solvents typically found in compound libraries.

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PolarScreen™ Nuclear Receptor Competitive Binding Assays Screening Data

PolarScreen™ Nuclear Receptor Competitive Binding Assays Screening Data

  
Figure 2. Competition data using the AR Competitor Assay, Green, generated on a 384-well plate. The concentration of the test compound that results in a half-maximal shift in polarization value equals the IC50 of the test compound, which is a measure of the relative affinity of the test compound for the androgen receptor ligand binding domain.
   
PolarScreen™ Nuclear Receptor Competitive Binding Assays Screening Data 
Figure 3. A subset of the Prestwick Bioactive Compound Library (bar code: 28151401), rich in nuclear receptor ligands was screened at 10 µM in the AR Competitor Assay, Green. Approximately 10% of the compounds were identified as hits because they displaced the Fluormone™ AL Green tracer from ARLBD (His-GST), and 1% of the library exhibited interfering compound fluorescence.

 

 

 

 

Screening Data

PolarScreen™ Nuclear Receptor Competitive Binding Assays Screening Data

PolarScreen™ Nuclear Receptor Competitive Binding Assays Screening Data

  
Figure 2. Competition data using the AR Competitor Assay, Green, generated on a 384-well plate. The concentration of the test compound that results in a half-maximal shift in polarization value equals the IC50 of the test compound, which is a measure of the relative affinity of the test compound for the androgen receptor ligand binding domain.
   
PolarScreen™ Nuclear Receptor Competitive Binding Assays Screening Data 
Figure 3. A subset of the Prestwick Bioactive Compound Library (bar code: 28151401), rich in nuclear receptor ligands was screened at 10 µM in the AR Competitor Assay, Green. Approximately 10% of the compounds were identified as hits because they displaced the Fluormone™ AL Green tracer from ARLBD (His-GST), and 1% of the library exhibited interfering compound fluorescence.

 

 

 

 

Resources

Resources

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

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