Products & Applications / Assay Ready Kits / Assay Ready Kit Format Selection Guide / Ab Hunter Bioassay Kit Assay Principle And How It Works

How It Works - Ab Hunter Bioassay Kit Principle

Ab Hunter™ Bioassay kits come with pre-validated ready-to-assay cells, detection reagents, ligand, cell plating reagent and assay plates. Simply thaw and plate cells, and run the assay with your antibodies. Assay-ready frozen cells are based on DiscoveRx's PathHunter® technology.

Assay Work Flow

Ab Hunter™ Bioassay Kits monitor the interaction of β-Arrestin with activated GPCRs (panel A). In this system, ProLink is appended to the C-terminus of the GPCR. Arrestin is fused to EA. Activation of the GPCR stimulates binding of Arrestin and forces complementation of the two enzyme fragments, resulting in a measurable chemiluminescent signal (panel B). Ab Hunter™ RTK Bioassay Kits measure the activation of the target receptor tyrosine kinase (panel C). Here, ProLink is fused to the receptor tyrosine kinase and EA is appended to SH2 phosphotyrosine domain protein. Receptor activation initiates tyrosine phosphorylation and recruitment of domain with SH2 protein resulting in active enzyme that can be detected with a chemiluminescent substrate (panel B).

A

B

C


AbHunter Assay Principle and work flow

Kit Components

  • Assay-ready, frozen PathHunter® cells
  • Optimized Cell Plating Reagent
  • Detection Reagents
  • Antibody Dilution Buffer
  • Control ligand
  • 96-well Tissue Culture Treated Plate