Rat Collagen Type IX Alpha 1 ELISA Kit

Manufactured for Antibodies.com, this sandwich Enzyme-Linked Immunosorbent Assay (ELISA) is designed for the in vitro quantitative determination of rat Collagen Type IX Alpha 1 (COL9A1) in tissue homogenates and other biological fluids. The assay employs the sandwich enzyme immunoassay technique, using an antibody specific for COL9A1 pre-coated onto a 96-well microplate. Standards and test samples are added to the wells, followed by a Biotinylated Anti-COL9A1 Antibody and an HRP-Avidin conjugate; a TMB substrate is then used to visualise the enzymatic reaction, producing a blue colour that turns yellow upon addition of the acidic stop solution. Absorbance is read at 450 nm in a microplate reader and the concentration of COL9A1 calculated against a standard curve.

The kit has a stated sensitivity of less than 0.52 ng/ml and a quantitative range of 1.56–100 ng/ml, with a total assay time of 4 hours 30 minutes. Detection is colorimetric, performed on a pre-coated microplate arranged as 12 x 8 well strips. Spike-and-recovery has been characterised in serum (average 91%, range 80–102%, n=5), EDTA plasma (average 91%, range 81–101%, n=5) and heparin plasma (average 84%, range 80–89%, n=5). Dilution linearity has been assessed at 1:2, 1:4, 1:8 and 1:16, giving ranges of 70–107%, 89–93%, 86–104% and 80–103% for serum; 95–100%, 85–110%, 81–99% and 90–102% for EDTA plasma; and 77–104%, 88–95%, 94–104% and 80–107% for heparin plasma.

Supplied components include a pre-coated 96-well microplate, lyophilised standard, detection solutions A and B, 30X wash buffer, sample dilution buffer, TMB substrate, stop solution and adhesive plate sealers, with individual storage temperatures ranging from +4°C to -20°C as indicated. Components should not be used past the stated expiration date.