Connective Tissue Growth Factor (CTGF), also known as CCN2 is a matricellular protein of the CCN family of extracellular matrix-ass°Ciated heparin-binding proteins. CTGF has important roles in many biological pr°Cesses, including cell adhesion, migration, proliferation, angiogenesis, skeletal development, and tissue wound repair, and is critically involved in fibrotic disease and several forms of cancers. Besides, Actin Beta (ACTb) has been identified as an interactor of CTGF, thus a binding ELISA assay was conducted to detect the interaction of recombinant rat CTGF and recombinant rat ACTb. Briefly, CTGF were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100uL were then transferred to ACTb-coated microtiter wells and incubated for 2h at 37°C. Wells were washed with PBST and incubated for 1h with anti-CTGF 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°C. Finally, add 50µL stop solution to the wells and read at 450nm immediately. The binding activity of of CTGF and ACTb was shown in Figure 1, and this effect was in a dose dependent manner.

(A) 3T3 cells were seeded into the well containting CTGF 1µg/mL and incubated for 1h at 37 °C; (B) 3T3 cells were seeded into the well without CTGF and incubated for 1h at 37°C.