Description
Principle of the assay: This assay employs a two-site sandwich ELISA to quantitative HYP in Rat serum, plasma. An antibody specific for HYP has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any HYP present is bound by the immobilized antibody. After removing any unbound substances, a biotin - conjugated antibody specific for HYP is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of HYP bound in the initial step. The color development is stopped and the intensity of the color is measured.
Background: 4-Hydroxyproline, or hydroxyproline (C5H9O3N), is an uncommon amino acid, abbreviated as HYP, e.g., in Protein Data Bank.Hydroxyproline differs from proline by the presence of a hydroxyl (OH) group attached to the C (gamma) atom. Other hydroxyprolines also exist in nature, notably 2, 3-cis 3, 4-trans-dihydroxyproline, which occurs in diatom cell walls, and is postulated to have a role in silica deposition. Hydroxyproline is also found in the walls of oomycetes, fungus-like protists related to diatoms. Hydroxyproline is produced by hydroxylation of the amino acid proline by the enzyme prolyl hydroxylase following protein synthesis (as a post-translational modification).Hydroxyproline is a major component of the protein collagen. Hydroxyproline and proline play key roles for collagen stability. They permit the sharp twisting of the collagen helix. It helps provide stability to the triple-helical structure of collagen by forming hydrogen bonds