This osteocalcin assay is a solid phase Enzyme Amplified Sensitivity Immunoassay performed on microtiter plates. The assay uses monoclonal antibodies (MAbs) directed against distinct epitopes of human osteocalcin. Calibrators and samples react with the capture monoclonal antibody (MAb 1) coated on microtiter well and with a monoclonal antibody (MAb 2) labelled with horseradish peroxidase (HRP). After an incubation period allowing the formation of a sandwich: coated MAb 1–human osteocalcin–MAb 2–HRP, the microtiter plate is washed to remove unbound enzyme labelled antibody. Bound enzymelabelled antibody is measured through a chromogenic reaction. Chromogenic solution (TMB ready for use) is added and incubated. The reaction is stopped with the addition of Stop Solution and the microtiter plate is then read at the appropriate wavelength. The amount of substrate turnover is determined colorimetrically by measuring the absorbance, which is proportional to the osteocalcin concentration. A calibration curve is plotted and OST concentration in samples is determined by interpolation from the calibration curve. The use of the EASIA reader (linearity up to 3 OD units) and a sophisticated data reduction method (polychromatic data reduction) result in a high sensitivity in the low range and in an extended calibration range.
E05 478 566 350 170 or Enzyme-Linked Immunosorbent Assays,E05 478 566 350 170 or Enzyme-Linked Immunosorbent Assays,Human proteins, cDNA and human recombinants are used in human reactive ELISA kits and to produce anti-human mono and polyclonal antibodies. Modern humans (Homo sapiens, primarily ssp. Homo sapiens sapiens). Depending on the epitopes used human ELISA kits can be cross reactive to many other species. Mainly analyzed are human serum, plasma, urine, saliva, human cell culture supernatants and biological samples.