Neuromedin U (NMU) is a neuropeptide hormone that is produced by neurons in the central and peripheral nervous systems. It is a member of the neuromedin family and is primarily found in the brain, where it is involved in pleiotropic physiological functions, including the regulation of blood pressure, food uptake, nociception, pain perception, bone formation, and immunological responses.Besides,G Protein Gamma 2 (GNg2) has been identified as an interactor of NMU, thus a functional binding ELISA assay was conducted to detect the interaction of recombinant human NMU and recombinant human GNg2. Briefly, biotin-linked NMU were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100μl were then transferred to GNg2-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST 3 times and incubation with Streptavidin-HRP for 30min, then wells were aspirated and washed 5 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 NMU and GNg2 was shown in Figure 1, the EC50 for this effect is 0.144ug/mL.