Active Adiponectin (ADPN) (APA605Ga01)

Human Adiponectin (ADPN) is a 53 kDa member of the PNPLA family of phospholipase A2 enzymes. It is a plasma protein secreted by adipose tissue showing pleiotropic effects with anti-diabetic, anti-atherogenic and anti-inflammatory properties. Adiponectin and AdipoR1 regulate PPARgC1a and mitochondria by Ca(2 ) and AMPK/SIRT1. Thus a functional binding ELISA assay was conducted to detect the interaction of recombinant chicken ADPN and recombinant rat PPARgC1a. Briefly, ADPN was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to PPARgC1a-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-ADPN 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 450/630 nm immediately. The binding activity of recombinant chicken ADPN and recombinant rat PPARgC1a was shown in Figure 1, the EC50 for this effect is 0.12 ug/mL.