Mouse Aggrecan ELISA Kit

From Antibodies.com, this sandwich Enzyme-Linked Immunosorbent Assay (sELISA) is designed for the in vitro quantitative determination of mouse Aggrecan in serum, plasma, tissue homogenates and other biological fluids. The procedure uses a pre-coated 96-well microplate (12 x 8 well strips) on which an antibody specific for Aggrecan is immobilised. Standards and samples are added so that the target binds to the captured antibody; after washing, a biotinylated anti-Aggrecan detection antibody is added, followed by HRP-Avidin conjugate and TMB substrate. The enzymatic reaction produces a blue product that turns yellow once the acidic stop solution is added, and absorbance is read at 450 nm in a colorimetric microplate reader, with the Aggrecan concentration calculated by reference to the standard curve.

According to the manufacturer's specifications, the assay has a sensitivity of <0.225 ng/ml and a standard curve range of 0.625-40 ng/ml, with a total assay time of 4 hours 30 minutes. In spike-and-recovery testing (n=5) values were 80-102% (average 91%) in serum, 81-100% (average 90%) in EDTA plasma and 80-89% (average 84%) in heparin plasma. Dilution linearity across 1:2 to 1:16 spanned 87-107% for serum, 79-108% for EDTA plasma and 82-105% for heparin plasma.

Kit components include a pre-coated 96-well microplate, two vials of lyophilised standard, detection solutions A and B (120 µl each), 30X wash buffer (20 ml), sample dilution buffer (45 ml), TMB substrate (9 ml), stop solution (6 ml) and five adhesive plate sealers. Components are stored at +4°C or -20°C as instructed in the protocol, and the reagents should not be used past the kit's expiration date.