SensoLyte® 520 FAP (Seprase) Assay Kit *Fluorimetric* - 100 Assays (96-well plate)

The 520 FAP Assay Kit is an assay for screening of enzyme inhibitors or for continuous assay of FAP activity. This kit utilizes a HiLyte Fluor™488/QXL® FRET substrate, that is cleaved by FAP, but not by DPPIV. Upon cleavage by FAP enzyme, the FRET substrate is separated into two fragments resulting in release of HiLyte Fluor™488 fluorescence which can be monitored at excitation/emission = 490 nm/520 nm. The long wavelength fluorescence of HiLyte Fluor™488 is less prone to interference by the autofluorescence of components in biological samples and test compounds, compared to shorter wavelength FRET substrates.

Fibroblast activation protein alpha or FAP, also known as Seprase, is an enzyme that belongs to the family of transmembrane serine proteases, and is most similar to DPPIV. In cancer, elevated FAP, in the tumor microenvironment is expressed by cancer associated fibroblasts (CAFs), that make up most of the tumor stroma and supports its development, progression, and invasion. FAP is also selectively expressed in stromal cells and mesenchymal stem cells during embryogenesis, wound healing, fibrotic reactions, and inflammatory conditions such as arthritis, atherosclerotic plaques, and fibrosis.

FAP has both dipeptidyl peptidase enzymatic activity and endopeptidase activity. DPPIV also has dipeptidylpetidase activity, but endopeptidase activity is specific to FAP and hence can be used to develop FAP specific detection methods.

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