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Products >  Cell_Biology >  Cell_Adhesion_and_Extracellular_Matrix >  EIA_Kits >  Heparan_Degrading_Enzyme_EIA_Kit

Heparan Degrading Enzyme ELISA Kit

Heparan sulfate is a complex and ubiquitous polysaccharide that is a component of mammalian cell membranes. It has been identified in mammalian liver, spleen, skin, placenta and platelets as well as in cancerous tissues such as melanomas, lymphomas, sarcomas, fibrosarcomas and colon cancer. Heparan sulfate has additionally been found to play an important defensive role in tumor cell invasion. The activity of heparan degrading enzyme (heparanase), an enzyme that cleaves heparan sulfate, is reported to be considerably higher in invasive cancer than normal cells. Thus, the correlation of heparan sulfate degrading enzyme activity with cancer malignancy is of much research interest.

Takara's Heparan Degrading Enzyme ELISA Kit provides a 96-well non-radioactive assay format for the measurement of heparan sulfate degradation in cultured cells, tissues or serum as well as for the screening of heparan sulfate degradation enzyme inhibitors. This kit is based on the bFGF(basic fibroblast growth factor)-binding ability of heparin-like molecules.

When heparan sulfate is degraded, it loses its ability to bind bFGF. Thus, the enzymatic activity of a sample can be quantitated by comparing the amount of bFGF-bound undegraded heparan sulfate with control heparanase-free total-bound heparan sulfate. A unique feature of Takara's Heparan Degrading Enzyme ELISA is the use of CBD-FGF, a fusion protein composed of the human fibronectin cell-binding domain and human fibroblast growth factor. CBD-FGF is immobilized on the surface of Takara's microtiter ELISA plates by an anti-fibronectin antibody that binds the CBD epitope. Termed DOC (Domain Oriented Capture), this solid phase attachment method enables the formation of a natural 3D bFGF structure for enhanced accessibility to and binding of heparan sulfate. Detection occurs via a colormetric assay method that uses biotinylated heparan sulfate as the substrate.

At-A-Glance Documents Images & Data Resources

Features

  • Safe: No radioactive isotopes required
  • 96-well format allows simultaneous assay of many samples
  • Any tissue, cell or serum, regardless of mammalian species, can be used.
  • Enables interaction studies of bFGF and heparin-like substances and other ligands
  • Unique solid phase attachment method creates a natural 3D bFGF structure for enhanced accessibility and binding to heparan sulfate

Applications

  • Quantitative measurement of heparan sulfate degradation in mammals
  • Screening of heparan sulfate degradation enzyme inhibitors
  • Heparan Degrading Enzyme ELISA

Components

Immobilized CBD-FGF microtiter plate

1 plate (96 wells)
Biotinylated heparan sulfate in reaction buffer 1 vial (for 5.5 mL)
Reaction buffer (for dilution)1 vial (11 mL)
Extraction buffer 1 vial (11 mL)
Standard (lyophilized)1 vial (for 250 mL)
Avidin-peroxidase conjugate (lyophilized) 1 vial (for 11 mL)
Peroxidase substrate 12 mL

Storage

2–8°C

Performance Characteristics

  • Range of assay: 0.1–4.2 U/mL
  • Assay sensitivity: 0.1 U/mL
  • Assay duration: 2-3 hours
  • Specificity: All animals
  • Test specimen type: Tissue, cultured cells, serum
  • Sample volume per well: 25 µL

    References

    1. Nakajima, M. et al. (1983) Nature 220:661-613.

    2. Nicolson, G. H. et al. (1998) Advan. Enzyme Regul. 38:19-32.

    3. Nakajima, M. et al. (1984) J. Biol. Chem. 259:2283-2290.

    4. Ricoveri, W. et al. (1986) Cancer Res. 46:3855-3861.

    5. Freeman, C. et al. (1997) Biochem. J. 325:229-237.

    6. Foxall, C. et al. (1995) Anal. Biochem. 231:366-373.

    7. Katayama, M. et al. (1989) Exp. Cell Res. 185:229-236.

    8. Hashi, H. et al. (1994) Cell Struct. Funct. 19:37-47.

    9. Nakajima, M. et al. (1991) J. Biol. Chem. 266:9661-9666.

    10. He, X. et al. (2004) Cancer Res. 64(11):3928-3933.




     
     
    Products
    Cat. # Product Package Size Price # of Units Select
    MK412 Heparan Degrading Enzyme Assay Kit 96 Assays $562.00
     

     



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