Wnt3a surrogate proteins address the challenge of producing native Wnt3a, which is notoriously difficult to express in active form due to extensive post-translational lipidation requirements. This full-length construct (aa 1–306) demonstrates robust canonical Wnt signaling activity in HEK293T cells, inducing Topflash reporter responses with an ED50 of 17.76–26.50 ng/mL—a potency range that supports dose-response studies in stem cell differentiation protocols and mechanistic investigations of β-catenin-dependent transcription. Mammalian expression ensures native-like glycosylation and folding, critical for receptor engagement in assays probing Frizzled–LRP5/6 interactions or screening small-molecule Wnt pathway modulators. The protein meets quality thresholds commonly required for cell-based functional assays, with purity exceeding 90% by SDS-PAGE and endotoxin below 1.0 EU/μg, making it appropriate for neuronal differentiation models, osteogenic lineage commitment studies, and receptor-ligand competition experiments where low background and consistent activity are essential.
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