Collagen Type XII (COL12), an extracellular matrix (ECM) protein, is a member of the fibrillar collagen family. COL12 is a minor collagen component found in various tissues, including cartilage, tendons, ligaments, blood vessel walls and corneas, it plays an important role in maintaining the structure and function of these tissues. It has been reported that the binding of COL12 and MMP12 is an important link in the remodeling and inflammation of ECM. Thus a functional binding ELISA assay was conducted to detect the interaction of recombinant human COL12 and recombinant mouse MMP12. Briefly, COL12 was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to MMP12-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-COL12 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 450/630 nm immediately. The binding activity of recombinant human COL12 and recombinant mouse MMP12 was shown in Figure 1, and this effect was in a dose dependent manner.