Anaspec has released the Sensolyte 520 Aggrecanase-1 Assay Kit, a fluorimetric assay optimised to detect AdamTS-4 (aggrecanase-1) activity.
It is a homogeneous Fret-based assay and has an emission in the green range (Ex/Em= 490 nm/520 nm).
The kit contains an 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.
AdamTS-4 (aggrecanase-1) is a major aggrecanase in human osteoarthritic cartilage.
Other members of the AdamTS family (AdamTS-1, -8, -9, -15 and - 16) have also been implicated in embryonic development and angiogenesis.
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 and an aggrecanase inhibitor.
