Milk Fat Globule EGF Factor 8 (MFGE8), also known as lactadherin, is a secreted glycoprotein containing EGF-like and coagulation factor C homology (C1/C2) domains. It mediates critical roles in apoptotic cell clearance (efferocytosis), tissue remodeling, angiogenesis, and immune regulation by binding integrin receptors (e.g., αvβ3/β5). MFGE8 facilitates phagocytosis of apoptotic cells via bridging phosphatidylserine on dying cells to phagocytes. It also modulates TGF-β activation and inflammatory responses. Expressed in macrophages, epithelial cells, and mammary glands, MFGE8 is implicated in atherosclerosis, autoimmune diseases, and cancer progression. Its dysfunction correlates with impaired tissue repair and aberrant immune activation.、Besides,EGF has been identified as an interactor of MFGE8, thus a functional binding ELISA assay was conducted to detect the interaction of recombinant mouse MFGE8 and recombinant human EGF. Briefly,MFGE8 was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to EGF-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-MFGE8 pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody for 1h at 37℃, wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50 μL stop solution to the wells and read at 450/630nm immediately. The binding activity of recombinant mouse MFGE8 and recombinant human EGF was shown in Figure 1, the EC50?for this effect is 0.043ug/mL.