Active Growth Differentiation Factor 15 (GDF15) (APC034Ra01)

Growth Differentiation Factor 15 (GDF-15), also called Macrophage Inhibitory Cytokine 1 (MIC-1), is a divergent member of the Transforming Growth Factor beta (TGF-beta) superfamily. Human GDF-15 shares 66% and 68% amino acid sequence identity with the rat and mouse proteins, respectively. GDF-15 is highly expressed in placenta and brain, and it is expressed at lower levels in kidney, pancreas, prostate, and colon. Similar to other TGF-beta family proteins, GDF-15 is synthesized as a large precursor protein that is cleaved at a dibasic cleavage site (RxxR) to release the mature protein. Biologically active GDF-15 is a disulfide-linked homodimer of the mature protein. GDF15 has been shown to have various functions, including inhibition of Tumor Necrosis Factor alpha (TNF-alpha) production from lipopolysaccharide-stimulated macrophages and the induction of cartilage formation. A functional binding ELISA assay was conducted to detect the interaction of recombinant rat GDF-15 and recombinant human GP9. Briefly, GDF-15 was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to GP9-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-GDF-15 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 GDF-15 and GP9 was shown in Figure 1, the EC50 for this effect is 0.47 ug/mL.