武漢云克隆科技股份有限公司

Fibronectin (FN) is a high-molecular weight (~440kDa) glycoprotein of the extracellular matrix that binds to membrane-spanning receptor proteins called integrins. Fibronectins bind cell surfaces and various compounds including collagen, fibrin, heparin, DNA, and actin. Fibronectin has numerous functions. For example, it involved in cell adhesion, cell motility, opsonization, wound healing, maintenance of cell shape, and so on. Besides, Lysyl Oxidase (LOX) has been identified as an interactor of FN, thus a binding ELISA assay was conducted to detect the interaction of recombinant mouse FN and recombinant human LOX. Briefly, LOX were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100ul were then transferred to FN-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-LOX pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 3 times. With the addition of substrate solution , wells were incubated 15-25 minutes at 37℃. Finally, add 50μL stop solution to the wells and read at 450nm immediately. Measured by its binding ability in a functional ELISA. When Recombinant FN is lmmobilized at 2 ug/mL(100 uLwell), the concentration of LOX that produces 50% optimal bindingresponse is found to be approximately0.03ug/mL.