Figure. The binding activity of FBN1 with FBLN2.
Fibrillin 1 (FBN1) is a 230-kb gene with 65 coding exons that encode a 2,871-amino-acid long proprotein called profibrillin which is proteolytically cleaved near its C-terminus by the enzyme furin convertase to give fibrillin-1, a member of the fibrillin family, and the 140-amino-acid long protein hormone asprosin. FBN1 is a large, extracellular matrix glycoprotein that serves as a structural component of 10-12nm calcium-binding microfibrils. These microfibrils provide force bearing structural support in elastic and nonelastic connective tissue throughout the body. Besides, Fibulin 2 (FBLN2) has been identified as an interactor of FBN1, thus a binding ELISA assay was conducted to detect the interaction of recombinant mouse FBN1 and recombinant mouse FBLN2. Briefly, FBN1 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100μL were then transferred to FBLN2-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-FBN1 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. The binding activity of FBN1 and FBLN2 was shown in Figure 1, and this effect was in a dose dependent manner.