Figure. The binding activity of MX1 with TUBb.
Interferon-induced GTP-binding protein Mx1 is a protein that in humans is encoded by the MX1 gene. In mouse, the interferon-inducible Mx protein is responsible for a specific antiviral state against influenza virus infection. The protein encoded by this gene is similar to the mouse protein as determined by its antigenic relatedness, induction conditions, physicochemical properties, and amino acid analysis. This cytoplasmic protein is a member of both the dynamin family and the family of large GTPases. Besides, Tubulin Beta (TUBb) has been identified as an interactor of MX1, thus a binding ELISA assay was conducted to detect the interaction of recombinant human MX1 and recombinant human TUBb. Briefly, MX1 was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100μL were then transferred to TUBb-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-MX1 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 MX1 and TUBb was shown in Figure 1, and this effect was in a dose dependent manner.