Cyt°Chrome P450 family 1 subfamily B member 1 (CYP1B1) is an enzyme, Which belongs to the cyt°Chrome P450 superfamily. CYP1B1 can catalyze many reactions involved in drug metabolism and synthesis of cholesterol, steroids, and other lipids. Besides, Heat Sh°Ck Protein family B (small) member 2 (HSPb2) has been identified as an interactor of CYP1B1, thus a binding ELISA assay was conducted to detect the interaction of recombinant human CYP1B1 and recombinant human HSPb2, CYP1B1 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100uL were then transferred to HSPb2-coated microtiter wells and incubated for 2h at 37°C. Wells were washed with PBST and incubated for 1h with anti-CYP1B1 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 CYP1B1 and HSPb2 was shown in Figure 1, and this effect was in a dose dependent manner.