武汉云克隆科技股份有限公司

Apolipoprotein A1 (APOA1) is the major protein component of HDL particles in plasma. It is a cofactor for lecithin cholesterolacyltransferase (LCAT) which is responsible for the formation of most plasma cholesteryl esters. ApoA1 was also isolated as a prostacyclin (PGI2) stabilizing factor, and thus may have an anticlotting effect. ApoA1 is often used as a biomarker for prediction of cardiovascular diseases. Besides, Plasminogen Activator, Urokinase (uPA) has been identified as an interactor of APOA1, thus a functional binding ELISA assay was conducted to detect the interaction of recombinant pig APOA1 and recombinant human uPA. Briefly, APOA1 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 ul were then transferred to uPA-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-APOA1 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 50 µL stop solution to the wells and read at 450/630 nm immediately. The binding activity of recombinant pig APOA1 and recombinant human uPA was shown in Figure 1, the EC50 for this effect was 0.03 ug/mL.