General Hydroxyproline ELISA Kit

Manufactured by Antibodies.com, this competitive enzyme-linked immunosorbent assay (cELISA) is designed for the in vitro quantitative determination of Hydroxyproline in serum, plasma, tissue homogenates or other biological fluids. The kit employs the competitive enzyme immunoassay technique using a pre-coated 96-well microplate arranged as 12 x 8 well strips. During incubation, Hydroxyproline present in samples or standards competes with a fixed amount of immobilised Hydroxyproline antigen for binding sites on a biotinylated anti-Hydroxyproline antibody. After incubation and washing, an HRP-avidin conjugate is added, followed by TMB substrate solution that produces a blue colour changing to yellow upon addition of acidic stop solution. The density of yellow is inversely proportional to the Hydroxyproline concentration, which is calculated by reading O.D. absorbance at 450 nm against a standard curve.

The assay has a product range of 78.1-5000 ng/ml and a sensitivity of <33 ng/ml. Total assay time is 2h 30m, with colourmetric detection performed on a colorimetric microplate reader. Reactivity is described as universal/general. Reported recovery values (n=5) are 82-102% (average 92%) for serum, 91-100% (average 95%) for EDTA plasma, and 88-100% (average 94%) for heparin plasma.

Kit components include a pre-coated microplate, lyophilised standard, Detection Solutions A and B, Wash Buffer (30X), Sample Dilution Buffer, TMB Substrate, Stop Solution, and plate sealers. Components should be stored at +4°C or -20°C as instructed, and the kit must not be used past its expiration date. The product has been cited in 1 publication.