Oncostatin M (OSM), encoded by the OSM gene, is a pleiotropic cytokine belonging to the interleukin-6 (IL-6) family. Initially identified for its anti-tumor activity, it is primarily secreted by activated T cells, macrophages, and dendritic cells. OSM plays pivotal roles in regulating cell growth, differentiation, inflammation, and tissue remodeling, with implications in immune responses, embryonic development, and pathological processes like fibrosis and autoimmune diseases. Its biological effects are mediated through binding to specific receptor complexes on target cell surfaces. OSM and LIF (Leukemia Inhibitory Factor) share structural homology and can bind to the common gp130 receptor subunit, enabling functional crosstalk in cellular signaling.Thus a functional ELISA assay was conducted to detect the interaction of recombinant mouse OSM and recombinant rat LIF.
Briefly, OSM was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to LIF-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-OSM 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 OSM and recombinant rat LIF was shown in Figure 1, the EC50?for this effect is 0.322μg/mL.