Wingless Type MMTV Integration Site Family, Member 5A (WNT5A) is a ligand for members of the frizzled family of seven transmembrane receptors. Can activate or inhibit canonical Wnt signaling, depending on receptor context. Stimulates cell migration. Decreases proliferation, migration, invasiveness and clonogenicity of carcinoma cells and may act as a tumor suppressor. Besides, WNT Inhibitory Factor 1 (WIF1) has been identified as an interactor of WNT5A, thus a binding ELISA assay was conducted to detect the interaction of recombinant mouse WNT5A and recombinant rat WIF1. Briefly, WNT5A were diluted serially in PBS, with 0.01%BSA (pH 7.4). Duplicate samples of 100uL were then transferred to WIF1-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-WNT5A 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 50µL stop solution to the wells and read at 450nm immediately. The binding activity of of WNT5A and WIF1 was shown in Figure 1, and this effect was in a dose dependent manner.