Figure. The binding activity of LpPLA2 with PTPRN.Phospholipase A2, Lipoprotein Associated (LpPLA2) also known as platelet-activating factor acetylhydrolase (PAF-AH) is a phospholipase A2 enzyme. LpPLA2 is platelet-activating factor (PAF) acetylhydrolase, a secreted enzyme that catalyzes the degradation of PAF to inactive products by hydrolysis of the acetyl group at the sn-2 position, producing the biologically inactive products LYSO-PAF and acetate. Besides, Protein Tyrosine Phosphatase Receptor Type N (PTPRN) has been identified as an interactor of LpPLA2, thus a binding ELISA assay was conducted to detect the interaction of recombinant human LpPLA2 and recombinant human PTPRN. Briefly, LpPLA2 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100uL were then transferred to PTPRN-coated microtiter wells and incubated for 2h at 37 degree C. Wells were washed with PBST and incubated for 1h with anti-LpPLA2 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 degree C. Finally, add 50uL stop solution to the wells and read at 450nm immediately. The binding activity of LpPLA2 and PTPRN was shown in Figure 1, and this effect was in a dose dependent manner.
SDS-PAGE
Western Blot
Sample: Recombinant LpPLA2, Human
Antibody: Rabbit Anti-Human LpPLA2 Ab (MBS2002593)
Phospholipase A2 Group VII (PLA2G7) which is also known as Lp-PLA2, is a plasma enzyme bound to lipoproteins: 80% bound to LDL, 15%-20% to HDL, and the remainder to VLDL. It is produced in major by mature macrophages and activated platelets and catalyzes the degradation of platelet-activating factor to biologically inactive products. Besides, Paraoxonase 1 (PON1) has been identified as an interactor of PLA2G7, thus a functional binding ELISA assay was conducted to detect the interaction of recombinant human PLA2G7 and recombinant rat PON1. Briefly, PLA2G7 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100 ul were then transferred to PON1-coated microtiter wells and incubated for 1h at 37?. Wells were washed with PBST and incubated for 1h with anti-PLA2G7 pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody, wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37?. Finally, add 50uL stop solution to the wells and read at 450/630 nm immediately. The binding activity of recombinant human PLA2G7 and recombinant rat PON1 was shown in Figure 1, the EC50 for this effect is 0.7 ug/mL
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