AXL Cellular Phosphorylation Assay (intracellular kinase activity assay) for compound screening and profiling in intact cells
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).
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.
Substrate phosphorylation as a readout of intracellular kinase activity via ELISA
More information can be found on our website Cellular Phosphorylation Assay Services.
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.