Description
Principle of the assay: human OPN ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for OPN has been precoated onto 96-well plates. Standards(NSO,I17-N300) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for OPN is added subsequently and then followed by washing with PBS or TBS buffer. Avidin-Biotin-Peroxidase Complex was added and unbound conjugates were washed away with PBS or TBS buffer. HRP substrate TMB was used to visualize HRP enzymatic reaction. TMB was catalyzed by HRP to produce a blue color product that changed into yellow after adding acidic stop solution. The density of yellow is proportional to the human OPN amount of sample captured in plate.
Background: Osteopontin (OPN) also called urinary stone protein, secreted phosphoprotein 1 (SPP1), bone sialoprotein, and early T lymphocyte activation 1 (ETA1). Osteopontin is a phosphorylated glycoprotein secreted to the mineralizing extracellular matrix by osteoblasts during bone development. It is believed to facilitate the attachment of osteoblasts and osteoclasts to the extracellular matrix, allowing them to perform their respective functions during osteogenesis.1Osteopontin is presumably involved in stone formation as stone matrix.2 The deduced protein sequence reveals a317-amino acid protein (34,982 Da) containing a 16-amino acid hydrophobic signal sequence and a 33,352-Da protein destines to undergo extensive post-translational modifications before being secreted from the cell. The gene is located on human chromosome 4.3 The standard product used in this kit is recombinant human OPN, consistingof 17-314 amino acid sequence with the molecular mass of 32.9KDa. As a result of glycosylation, the molecular mass is 60-65KDa