Detection Method:Sandwich
Test principle:This assay employs a two-site sandwich ELISA to quantitate ZBED3 in samples. An antibody specific for ZBED3 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyZBED3 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for ZBED3 is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of ZBED3 bound in the initial step. The color development is stopped and the intensity of the color is measured.
Product Overview:Zbed3 contains a PPPPSPT motif, which is crucial to its binding to Axin. The Ser and Thr residues in the motif appear to be also phosphorylated by glycogen synthase kinase 3β (GSK3β) and the CKI family kinases, as GSK3β and CKI? could enhance the interaction of Zbed3 with Axin. Expressing Zbed3, but not these mutants, led to inhibition of GSK3β-mediated β-catenin phosphorylation, cytoplasmic β-catenin accumulation, and activation of lymphoid enhancer binding factor-1-dependent reporter gene transcription. Furthermore, knockdown of Zbed3 with RNA interference attenuated Wnt-induced β-catenin accumulation, lymphoid enhancer binding factor-1-dependent luciferase reporter activity, and the Wnt target gene expression. Zbed3 is a novel Axin-binding protein that is involved in Wnt/β-catenin signaling modulation.
