Aggrecanase Sensitive Activity Assay
The Aggrecanase Sensitive Activity Assay is For Research Use Only
Size: 1×96 wells
Dynamic Range: 0.56 – 100 pM
Incubation Time: 4 hours
Sample Type: cell culture
Sample Size: 5 µL
ARGSVIL-peptide-s standard, proteolytic digested aggrecan-IGD-s with standard aggrecanase and test samples are incubated in microtiter wells pre-coated with anti-ARGSVIL-neoepitope antibody. ARGSVIL-peptide-s is bound to the coated antibody, while other components are removed with washing and aspiration. The bound ARGSVIL-peptide-s is detected with a second peroxidase-labeled antibody. Any excess of the conjugate is removed with washing and aspiration. The amounts of peroxidase bound to different wells are determined in reactions with peroxidase substrate TMB. The reactions are stopped by addition of sulfuric acid solution and absorbance is read at 450 nm in a microtiter plate spectrophotometer.
The assay kit consists of two modules, the Aggrecanase module and the ELISA Module. The modified aggrecan interglobular domain (aggrecan-IGD-s) is first digested with Aggrecanase (ADAMTS4). Proteolytic cleavage of the substrate releases an aggrecan peptide with the Nterminal sequence ARGSVIL (ARGSVIL-peptide-s). The ARGSVIL-peptide-s is then quantified with two monoclonal anti-peptide antibodies.