Figure. The binding activity of MIF with MHCDG.
Macrophage migration inhibitory factor (MIF), also known as glycosylation-inhibiting factor (GIF), L-dopachrome isomerase, or phenylpyruvate tautomerase is a protein classified as an inflammatory cytokine. MIF is an important regulator of innate immunity. It involved in cell-mediated immunity, immunoregulation, and inflammation. MIF plays a role in the regulation of macrophage function in host defense through the suppression of anti-inflammatory effects of glucocorticoids. This lymphokine and the JAB1 protein form a complex in the cytosol near the peripheral plasma membrane, which may indicate a role in integrin signaling pathways. Besides, Major Histocompatibility Complex Class II Invariant Chain (MHCDG) has been identified as an interactor of MIF, thus a binding ELISA assay was conducted to detect the interaction of recombinant human MIF and recombinant human MHCDG. Briefly, MIF were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100uL were then transferred to MHCDG-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-MIF 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 MIF and MHCDG was shown in Figure 1, and this effect was in a dose dependent manner.