Myostatin (MSTN) also known as growth differentiation factor 8 (GDF-8) is a member of the TGF beta protein family. Myostatin is a secreted growth differentiation factor that produced and released by myocytes. This protein negatively regulates skeletal muscle cell proliferation and differentiation. Besides, Bone Morphogenetic Protein 1 (BMP1) has been identified as an interactor of MSTN, thus a binding ELISA assay was conducted to detect the interaction of recombinant human MSTN and recombinant human BMP1. Briefly, MSTN were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100uL were then transferred to BMP1-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-MSTN 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 MSTN and BMP1 was shown in Figure 1, and this effect was in a dose dependent manner.
Figure. The binding activity of MSTN with BMP1.