Description
Principle of the assay: human Seprase ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from mouse specific for Seprase has been precoated onto 96-well plates. Standards(NSO, L26-D760) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for Seprase 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 Seprase amount of sample captured in plate.
Background: FAP (Fibroblast Activation Protein, Alpha) also known as FAPA or SEPRASE, is an inducible cell surface glycoprotein that was originally identified in cultured fibroblasts using monoclonal antibody F19. The protein encoded by this gene is a homodimeric integral membrane gelatinase belonging to the serine protease family. The FAP gene is mapped on 2q24.2. FAP is most closely related to DPPIV and they share about 50% of their amino acids. FAP is catalytically active as a 170kD dimer and has dipeptidase and gelatinase activity. Its gelatinase activity requires a glycine in P2 position.FAP-alpha shows 48% amino acid identity with dipeptidyl peptidase IV and 30% identity with DPP4-related protein. Northern blot analysis detected a 2.8-kb FAP-alpha mRNA in fibroblasts. Depletion of FAP-expressing cells, which made up only 2% of all tumor cells in established Lewis lung carcinomas, caused rapid hypoxic necrosis of both cancer and stromal cells in immunogenic tumors by a process involving interferon-gamma and tumor necrosis factor-alpha