AXL Cellular Phosphorylation Assay Service

AXL Cellular Phosphorylation Assay (intracellular kinase activity assay) for compound screening and profiling in intact cells

Target Overview

Target Description

AXL is a receptor tyrosine kinase with a unique structure of the extracellular region that juxtaposes IgL and FNIII repeats. AXL stimulation has been described to drive cellular proliferation and the protein growth arrest-specific gene-6 (GAS6) has been found to be a ligand. Dysregulation of AXL and its ligand GAS6 is implicated in the pathogenesis of several human cancers. Recently, it has been shown by siRNA knock-down that AXL plays a role in tumor growth, metastasis and angiogenesis (Li, Y et al. (2009) Oncogene).

HGNC Reference

AXL

Synonyms

EC, UFO

Cell Line

MEF

Target Expression

Transfected

Assay Properties

Assay Description

Reaction Biology’s cellular AXL phosphorylation assay was generated on a mouse embryonal fibroblast (MEF) background. Cells were transfected to express a full-length AXL protein. After clonal selection a transformed cell line with a high level of autophosphorylated AXL was obtained. By adding Sunitinib phospho-AXL levels are largely decreased and thus the dynamic behaviour to determine inhibitory potentials of compounds was achieved. Phospho-AXL levels are quantified by Sandwich-ELISA technique.

Readout

Substrate phosphorylation as a readout of intracellular kinase activity via ELISA

Screening Location

Freiburg, Germany

Further Information

More information can be found on our website Cellular Phosphorylation Assay Services.

Reference compound IC50 for AXL

Reference compound IC50 for AXL

Sunitinib is a potent inhibitor of the phospho-AXL signal found in the described cells. The graph shows a representative result.