Active Phospholipase A2, Group IIA (PLA2G2A)

Phospholipase A2, Group IIA (PLA2G2A) is a secreted calcium-dependent enzyme that hydrolyzes phospholipids at the sn-2 position to release arachidonic acid and lysophospholipids, key mediators of inflammation. Primarily expressed in inflammatory cells and various tissues, PLA2G2A plays critical roles in host defense, lipid metabolism, and inflammatory diseases like atherosclerosis and arthritis. Its overexpression is associated with chronic inflammatory conditions and cancer progression. PLA2G2A interacts with Phospholipase A2 Group VII (PLA2G7) by coordinating lipid hydrolysis pathways, where PLA2G7's lipoprotein-associated activity modulates PLA2G2A's pro-inflammatory effects in vascular and immune contexts.Thus a functional binding ELISA assay was conducted to detect the interaction of recombinant human PLA2G2A and recombinant human PLA2G7. Briefly, PLA2G2A was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to PLA2G7-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-PLA2G2A 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 450/630nm immediately. The binding activity of recombinant human PLA2G2A and recombinant human PLA2G7 was shown in Figure 1, the EC50 for this effect is 0.038 µg/mL.