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This kit captures ADAMTS13-cleaved products using a sandwich method with the anti-GST mouse monoclonal antibody and the peroxidase-conjugated mouse monoclonal anti-N10 antibody. First, the anti-GST mouse monoclonal antibody immobilized on to the microplate reacts with the VWF73 substrate（GST-VWF73-His）. The sample is then applied to the microplate, on which ADAMTS13 cleaves the VWF73 substrate. By applying the mouse monoclonal anti-N10 antibody conjugated with horseradish peroxidase（HRP conjugated antibody）, the cleavage product is sandwiched between the two antibodies. Because the amount of the cleavage products depends on the ADAMTS13 activity, the amount of the enzyme-labeled antibodies that bind to the cleavage products also reflect the level of ADAMTS13 activity. Therefore, the plasma ADAMTS13 activity can be determined by colorimetrically measuring the amount of detached oxidized （colored）TMBZ using urea hydrogen peroxide（H2O2）and 3,3’,5,5’-Tetramethyl- benzidine（TMBZ）as the substrates. |
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