AnaSpec Introduces Industry’s First Fluorimetric Assay Kit for Aggrecanase-1 Activity Detection

AnaSpec has announced the release of the industry’s first fluorimetric assay optimized to detect ADAMTS-4 (aggrecanase-1) activity - the SensoLyteTM 520 Aggrecanase-1 Assay Kit. A FRET based assay, it is homogeneous and has an emission in the green range (Ex/Em= 490 nm/520 nm).

The kit contains a novel internally quenched peptide substrate for aggrecanase-1 consisting of 5-FAM/TAMRA as the FRET pair. In the intact FRET peptide, the fluorescence of 5-FAM is quenched by TAMRA. Active ADAMTS4 cleaves FRET substrate into two separate fragments resulting in an increase of 5-FAM fluorescence. Other members of aggrecanase family, such as ADAMTS-1 and ADAMTS-5, do not cleave the substrate provided in this kit.

Aggrecanases are members of the ADAMTS (A Disintegrin And Metalloprotease with Thrombospondin motif) family of proteases. ADAMTS-4 (Aggrecanase-1) and ADAMTS-5 (Aggrecanase-2) activities result in the degradation of cartilage aggrecan in arthritic diseases (1-3). ADAMTS-4 (aggrecanase-1) is a major aggrecanase in human osteoarthritic cartilage (4). Other members of the ADAMTS family (ADAMTS-1, -8, -9, -15 and –16) have also been implicated in embryonic development and angiogenesis (5).

The kit provides enough reagents to perform 100 assays in a 96-well format.

The kit contains:
• Aggrecanase-1 FRET substrate
• 5-FAM reference standard for calibration
• Assay buffer
• Aggrecanase inhibitor
• Stop Solution.

References :
1. Gur, PK. et al. J. Clin. Invest. 1059, 1335 (2000).
2. Tortorella, MD. Science 284, 1663 (1999).
3. Bonderson, J. Clin. Exp. Rheumatol. 26, 139 (2008).
4. Naito, S, et al. Pathol. Int. 57, 703 (2007).
5. Tang, BL. Int. J. Biochem. Cell Biol. 33, 33 (2001).