PathHunter® Nuclear Hormone Receptor Protein Translocation Assay Platform

PathHunter nuclear translocation  assay detects binding of an agonist or an antagonist to the ProLabel-tagged NHR of interest leading to the translocation of NHR to nucleus. The complementation occurs between the two β-galactosidase components: the ProLabel tag which is fused to the NHR of interest; the Enzyme Acceptor (EA) is sequestered into nuclear compartment. These components interact once they are in the same nuclear compartment, forming active β-gal enzyme that converts substrate to detectable signal.


Technology Principle

Pathhunter Nuclear Hormone TranslocationPlatform Schematic
 

Measure specific translocation of ProLabel tagged Glucocorticoid into the nucleus

No Dex  

With Dex

  

Dex Treated GR NHRNT Cells

 
Immunofluorescence analysis of Dexamethasone induced Glucocorticoid Receptor translocation into the nucleus using PathHunter NHRNT cells.
 
PathHunter GR NHRNT cells were plated in a 384-well plate at 10,000 cells/well and stimulated with Dexamethasone for 3 hours. Signal was detected using the PathHunter® Detection Kit (93-0001) according to the recommended protocol.

Features & Benefits

  • Fewer off-target effects, due to shorter assay time (6 hours)
  • Measures NHR activation and translocation
  • Employs full length NHRs, providing more biologically relevant data
  • Applicable to high-throughput data
  • Simple plate-treat-read format