Human Collagen Type II Alpha 1 ELISA Kit

From Antibodies.com, this Sandwich ELISA kit is designed for the in vitro quantitative determination of human Collagen Type II Alpha 1 in serum, plasma or other biological fluids. Detection is read on a colorimetric microplate reader at 450 nm using a pre-coated 96-well microplate arranged as 12 x 8 well strips.

The kit employs the sandwich enzyme immunoassay technique. Standards and samples are added to the wells, where the target is captured by an immobilized antibody. After washing, a Biotinylated Anti-Collagen II Antibody is added, followed by an HRP-Avidin conjugate. TMB substrate is then used to visualise the enzymatic reaction, producing a blue product that turns yellow upon addition of the acidic stop solution; the absorbance at 450 nm is proportional to the amount of Collagen II captured, and concentration is calculated by reference to a standard curve. Kit components are stored at +4°C or -20°C as instructed.

Reported sensitivity is <0.227 ng/ml and the assay range is 0.625–40 ng/ml, with a total assay time of 4 hours 30 minutes. Recovery in serum (n=5) ranged from 80% to 102% (average 91%); in EDTA plasma (n=5) from 81% to 101% (average 91%); and in heparin plasma (n=5) from 80% to 89% (average 84%). Linearity across dilutions 1:2, 1:4, 1:8 and 1:16 was reported as follows: serum (n=5) 70–107%, 89–93%, 86–104%, 80–103%; EDTA plasma (n=5) 95–100%, 85–110%, 81–99%, 90–102%; heparin plasma (n=5) 77–104%, 88–95%, 94–104%, 80–107%.