FAK Cellular Phosphorylation Assay Service

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

Target Overview

Target Description

The focal adhesion kinase (FAK) is a non-receptor tyrosine kinase that plays an important role in signal transduction pathways of integrin or growth factor receptor mediated cell adhesion. FAK has an important role in a large number of processes involved in the development and progression of cancer (survival, migration, proliferation and invasion). FAK overexpression/activity is found in a variety of human cancers making it an attractive target for anti-cancer therapy.

HGNC Reference

PTK2

Synonyms

FAK, FADK

Cell Line

MEF

Target Expression

Transfected

Assay Properties

Assay Description

The cellular phosphorylation assay for FAK determines the autophosphorylation on Tyr397 of the FAK protein. Murine embryonal fibroblast cells deficient of the endogenous FAK express a human full-length FAK which is constantly phosphorylated in adherent cells. By adding Staurosporine phospho-FAK-Tyr397 levels are largely decreased and thus the dynamic behaviour to quantify inhibitory potentials of compounds can be obtained. The assay is validated based on known inhibitors of FAK kinase activity (see Fig. 1).

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 FAK

Reference compound IC50 for FAK

Staurosporine is an inhibitor of the phospho-FAKTyr397 signal found in the described cells. The graph shows a representative result.