武漢云克隆科技股份有限公司

Wingless Type MMTV Integration Site Family, Member 10A (WNT10A) is a ligand for members of the frizzled family of seven transmembrane receptors. It is involved in the regulation of cell proliferation, differentiation, migration, and apoptosis. Overexpression of WNT10A has been associated with various diseases, including cancer, where it can contribute to tumorigenesis through activation of the Wnt-beta-catenin-TCF signaling pathway. LRP5 and LRP6, as co-receptors of Wnt signal, play a key role in the signal transduction process, thus a functional binding ELISA assay was conducted to detect the interaction of recombinant mouse WNT10A and recombinant rat LRP5. Briefly, WNT10A was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to LRP5-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-WNT10A 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 mouse WNT10A and recombinant rat LRP5 was shown in Figure 1, the EC50?for this effect is 0.026 ug/mL.