Takara's Heparan Degrading Enzyme Assay Kit provides a 96-well non-radioactive assay for the measurement of heparan degrading enzyme activity in cultured cells, tissues, or serum as well as screening of heparan sulfate degrading enzyme inhibitors. This kit is based upon the property of heparin-like molecules to bind bFGF (basic fibroblast growth factor).
When heparan sulfate is degraded by heparan sulfate degrading enzyme, it loses its ability to bind to bFGF. Thus, the enzymatic activity of a sample can be quantitated by comparing the amount of bFGF-bound undegraded heparan sulfate of the sample to total-bound heparan sulfate of a heparanase-free control. A unique feature of this kit is the use of CBD-FGF, a fusion protein composed of human fibronectin cell-binding domain and human fibroblast growth factor. CBD-FGF is immobilized on the surface of Takara's microtiter plate by an anti-fibronectin antibody containing an epitope in the CBD region. Termed DOC (Domain Oriented Capture), this solid phase attachment method allows a natural 3-dimensional bFGF structure for enhanced accessibility and binding to heparan sulfate. Detection is by a colormetric method which uses biotinylated heparan sulfate as the substrate.
Heparan sulfate is a ubiquitous complex polysaccharide which is present as a component of mammalian cell membranes. It has been identified in mammalian liver, spleen, skin, placenta, and platelets, as well as in cancerous tissue such as melanomas, lymphomas, sarcomas, fibrosarcomas, and colon cancer. Furthermore, heparan sulfate has been shown to play an important defensive role against invasion of tumor cells. The activity of heparan sulfate degrading enzyme (or heparanese), an enzyme that cleaves heparan sulfate, has been reported to be considerably higher in invasive cancer cells than in normal cells. Thus, the correlation of heparan sulfate degrading enzyme activity with malignancy of cancer cells has gained much interest.
