Intended use
This immunoassay kit allows for the specific measurement of rat OT concentrations in serum and plasma.
Introduction
Osteocalcin (OT) is the most abundant noncollagenous protein in mature bone where it constitutes 1% to 2% of the total protein. Synthesized by osteoblasts, it is incorporated into the bone matrix. Human osteocalcin is a 49-residue protein (MW. 5669) with three gamma-carboxyglutamic acid residues, at positions 17, 21 and 24. These three residues confer on it a very strong ability to bind to hydroxyapatite.
Some of the newly synthesized osteocalcin escapes into the blood and its concentration in serum reflects the rate of bone formation rather than the rate of bone resorption. Although its function is not known, it is a more specific biochemical index of bone metabolism than other more established indices such as urinary hydroxyproline and total serum alkaline phosphatase activity.
Test principle
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal
antibody specific for OT has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any OT present is bound by the immobilized antibody. An enzyme-linked monoclonal antibody specific for OT is added to the wells. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of OT bound in the initial step. The color development is stopped and the intensity of the color is measured.
