Figure. The binding activity of IGF1 with IGFBP3.
Insulin-like growth factor 1 (IGF1), also called somatomedin C is a hormone similar in molecular structure to insulin. It plays an important role in childhood growth and continues to have anabolic effects in adults. A synthetic analog of IGF1, mecasermin, is used for the treatment of growth failure. Besides, Insulin Like Growth Factor Binding Protein 3 (IGFBP3) has been identified as an interactor of IGF1, thus a binding ELISA assay was conducted to detect the interaction of recombinant mouse IGF1 and recombinant mouse IGFBP3. Briefly, IGF1 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100μL were then transferred to IGFBP3-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-IGF1 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 IGF1 and IGFBP3 was shown in Figure 1, and this effect was in a dose dependent manner.