Wingless-type MMTV integration site family, member 16 is a protein that in humans is encoded by the WNT16 gene. The WNT gene family consists of structurally related genes that encode secreted signaling proteins. These proteins have been implicated in oncogenesis and in several developmental processes, including regulation of cell fate and patterning during embryogenesis. This gene is a member of the WNT gene family. It contains two transcript variants diverging at the 5' termini. These two variants are proposed to be the products of separate promoters and not to be splice variants from a single promoter. Besides, Tubulin Beta 3 (TUBb3) has been identified as an interactor of WNT16, thus a binding ELISA assay was conducted to detect the interaction of recombinant human WNT16 and recombinant human TUBb3. Briefly, WNT16 were diluted serially in PBS, with 0.01% BSA (pH 7.4). Duplicate samples of 100μL were then transferred to TUBb3-coated microtiter wells and incubated for 2h at 37℃. Wells were washed with PBST and incubated for 1h with anti-WNT16 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 WNT16 and TUBb3 was shown in Figure 1, and this effect was in a dose dependent manner.
Figure. The binding activity of WNT16 with TUBb3.