Figure. The binding activity of TIMP3 with MMP2.
Tissue Inhibitors Of Metalloproteinase 3 (TIMP3) is an protein belongs to the tissue inhibitor of metalloproteinases family. They are inhibitors of the matrix metalloproteinases. TIMP-3 is the only member of the TIMP family which is found exclusively in the extracellular matrix (ECM). It is regulated in a cell cycle-dependent fashion in certain cell types and may serve as a marker for terminal differentiation. Besides, Matrix Metalloproteinase 2 (MMP2) has been identified as an interactor of TIMP3, thus a binding ELISA assay was conducted to detect the interaction of recombinant mouse TIMP3 and recombinant mouse MMP2. Briefly, TIMP3 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100µl were then transferred to MMP2-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-TIMP3 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 TIMP3 and MMP2 was shown in Figure 1, and this effect was in a dose dependent manner.