Active Paraoxonase 1 (PON1)

Paraoxonase 1 (PON1) is responsible for hydrolysing organophosphate pesticides and nerve gasses. PON1 (paraoxonase 1) is also a major anti-atherosclerotic component of high-density lipoprotein (HDL). Besides, Clusterin (CLU) has been identified as an interactor of PON1, thus a binding ELISA assay was conducted to detect the interaction of recombinant human PON1 and recombinant human CLU. Briefly, PON1 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100uL were then transferred to CLU-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-PON1 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 of PON1 and CLU was shown in Figure 1, and this effect was in a dose dependent manner.