Description
Principle of the assay: mouse MFGE8 ELISA Kit was based on standard sandwich enzyme-linked immune-sorbent assay technology. A monoclonal antibody from rat specific for MFGE8 has been precoated onto 96-well plates. Standards(NSO, A23-C463) and test samples are added to the wells, a biotinylated detection polyclonal antibody from goat specific for MFGE8 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 mouse MFGE8 amount of sample captured in plate.
Background: MFGE8 (Milk Fat Globule-Egf Factor 8), also called as Lactadherin or SED1, is a protein which in humans is encoded by the MFGE8 gene. Mfge8 is secreted protein found in vertebrates, including mammals as well as birds. By fluorescence in situ hybridization, the MFGE8 gene was mapped to chromosome 15q25. MFGE8 is a factor that links apoptotic cells to phagocytes. It specifically bound to apoptotic cells by recognizing aminophospholipids such as phosphatidylserine. MFGE8, when engaged by phospholipids, bound to cells via its RGD motif. It bound particularly strongly to cells expressing alpha-V-beta-3 integrin. It showed that Mfge8 was expressed in intestinal lamina propria macrophages in mice. Using a wound-healing assay, they showed that Mfge8 promoted migration of intestinal epithelial cells through a PKC-epsilon (PRKCE)-dependent mechanism