FGFR2 Kinase Inhibitor Screening Assay Kit

Intended Use

The FGFR2 Kinase Inhibitor Screening Assay Kit is designed to measure the activities of recombinant catalytic domain of FGFR2 (Fibroblast Growth Factor Receptor 2) for the rapid and sensitive evaluation of inhibitors or activators. The phosphotyrosine specific monoclonal antibody used in this assay kit has been demonstrated to recognize the phosphotyrosine residue in recombinant "Tyrosine kinase-substrate-1", which is efficiently phosphorylated by FGFR2 in vitro.

Applications of this kit include:
1) Screening inhibitors or activators of recombinant catalytic domain of FGFR2.
2) Detecting the effects of pharmacological agents on recombinant catalytic domain of FGFR2.

This assay kit is for research use only and not for use in diagnostic or therapeutic procedures.

Storage

Upon receipt store all components at 4°C.
Don't expose reagents to excessive light.

Performance Characteristics

The FGFR2 Kinase Inhibitor Screening Assay Kit has been shown to detect the kinase activity of recombinant catalytic domain of FGFR2. The assay shows good linearity of sample response.

General Description

The fibroblast growth factors (FGFs) belong to a family of about 23 related polypeptides. They display biological activity toward cells of mesenchymal, neuronal, and epithelial origin and are involved in processes such as cell growth, organ development, and angiogenesis. The biological effects of FGFs are exerted through interactions with FGF receptors (FGFRs). The receptor family consists of four known members, FGFR1-4, with many isoforms. Upon ligand binding the receptor is thought to be activated through dimerization and phosphorylation by the intracellular tyrosine kinase domains. Heparan sulfate proteoglycans (HSPGs), abundant components of cell surfaces and the extracellular matrix, appear central to signaling through FGF·FGFR complexes.
Overexpression and amplification of growth factor receptors are common alterations observed in human breast cancer cells. The FGFR1 and FGFR2 genes are expressed in both normal and breast cancer tissues, and alterations, including amplification and overexpression of FGFR1 and FGFR2, have previously been reported in 5-10% of primary breast cancer specimens.
In this decade, because of the localization of FGFR2 to the same chromosomal region as the mutation responsible for Crouzon syndrome, FGFR2 became a candidate gene for the clinical disorder. It has been demonstrated that the mutations in FGFR2 gene was found in patients with Crouzon syndrome, Apert syndrome, Pfeiffer syndrome, and Jackson-Weiss syndrome.

Measurement of FGFR2 Kinase activity

The protocol generally regarded as most sensitive for the quantitative measurement of FGFR2 kinase activity involves incubation of the FGFR2 kinase sample with substrate, either a natural or synthetic polypeptide (such as poly[Glu,Tyr]4:1), in the presence of Mg2+, Mn2+ and 32P-labeled ATP. The reaction is terminated by "spotting" a sample onto a filter paper disc, followed by immersion in acid to precipitate the radiolabeled product. The filter papers are then washed extensively to remove unincorporated radiolabel and the radioactivity is counted. While sensitive, this method is labor-intensive, generates hazardous radioactive waste, and depends on a radioisotope of short half-life. It is particularly unsuitable when kinase assays are only performed on an infrequent basis. The Research Product FGFR2 Kinase Inhibitor Screening Assay Kit uses a horseradish peroxidase coupled anti-phosphotyrosine monoclonal antibody as a reporter molecule in a 96-wells ELISA format. This assay provides a non-isotopic, sensitive and specific method to detect kinase activity of recombinant FGFR2 catalytic domain.