Monokine Induced By Interferon Gamma (MIg) is a pro-inflammatory cytokine primarily secreted by immune cells like macrophages and T cells upon IFN-γ stimulation. It plays pivotal roles in regulating immune responses, promoting leukocyte recruitment, and mediating anti-tumor and anti-pathogen defenses. Dysregulation of MIg is linked to autoimmune diseases and chronic inflammation. MIg binds to CXCL12 (SDF1, A122Hu) with moderate affinity, forming a complex that modulates chemotaxis and immune cell trafficking.Thus a functional ELISA assay was conducted to detect the interaction of recombinant human Mlg and recombinant human SDF1. Briefly, Mlg was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to SDF1-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-Mlg 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 human Mlg and recombinant human SDF1 was shown in Figure 1, the EC50?for this effect is 2.50 μg/mL.