Products for Wnt Research

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Wnt Signaling Pathways

Wnt ligands are a large family of secreted glycoproteins that have a central role in many processes involved in embryonic development and adult tissue homeostasis. The biological effects of Wnt ligands are mediated through an increasingly complex interplay of Wnt receptors, transmembrane regulators, and soluble inhibitors. The combination of these proteins are critical in determining whether a particular Wnt ligand will initiate b-Catenin-dependent or

b-Catenin-independent signaling cascades. R&D Systems is globally recognized as providing the highest quality and largest selection of Wnt proteins on the market. This brochure highlights our breadth of Wnt-related reagents including difficult-to-isolate Wnt ligands, our selection of recently discovered transmembrane regulators of Wnt (RNF43, ZNRF3, LGR 4-6, and TROY), our range of Wnt-related antibodies, and Tocris small molecules that target the Wnt pathway.

Proteins for Wnt Research

Wnt Ligands: Most Extensive Offering!

Recombinant Mouse Norrin Activates b-Catenin/TCF Through Frizzled-4 and LRP-5. The b-catenin/TCR reporter activity (RFU) in HEK293 human embryonic kidney cells overexpressing human Frizzled-4 and human LRP-5 was dose-dependently increased by Recombinant Mouse (rm)Norrin (Catalog # 3497-NR). In this assay the typical ED50 for rmNorrin ranges from 40–200 ng/mL.

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Recombinant Mouse Wnt-8a Activates Osteoblast Differentiation of Mouse Mesenchymal Stem Cells. Addition of Recombinant Mouse (rm)Wnt-8a (Catalog # 8419-WN) to C3H10T1/2 mouse mesenchymal stem cells induced osteoblast differentiation as quantified with a dose responsive increase in alkaline phosphatase production. In this assay the typical ED50 rmWnt-8a ranges from 0.5–2.5 µg/mL.

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Regulators of Wnt Signaling: New and Emerging Proteins

Robust Binding Activity for Recombinant Human ZNRF3 and RNF43. Binding activity of Recombinant Human (rh)ZNRF3 and rhRNF43 for rhR-Spondin 3 (Catalog # 4575-RS) was analyzed using ELISA. rhZNRF3 binds to rhR-Spondin 3 with a Kd = 0.2 nM (blue line). rhRNF43 binds to rhR-Spondin 3 with a Kd = 0.1 nM.

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Cutting Edge Research from R&D Systems

R&D Systems takes pride in being the leading manufacturer of Wnt family proteins as well as an incubator for scientists to perform cutting edge research to advance our knowledge of Wnt signaling.

Lgr4–6 Show Different R-Spondin Binding Preferences and Affinities

Analysis of Lgr Receptor Affinity for R-Spondin. Binding Activity of Recombinant Human (rh)Lgr4, rhLgr5, and rhLgr6 to rhR-Spondin 1 (blue), rhR-Spondin 2 (red), rhR-Spondin 3 (green), and rhR-Spondin 4 (peach) was analyzed using ELISA. (A) Binding curves of rhLgr4 with rhR-Spondins 1–4. (B) Binding curves of Lgr5 with rhR-Spondins 1–4. (C) Binding curves of Lgr6 with rhR-Spondins 1–4. (D) Table of dissociation constants (Kd) for all Lgr/R-Spondin combinations tested.

Antibodies for Wnt Research

Lgr5/GPR49 Antibodies: Qualified by Application

Detection of Lgr5/GPR49 by Flow Cytometry. The HEK293 human embryonic kidney cell line was transfected with eGFP-tagged mouse Lgr5/GPR49 and stained with either (A) Rat Anti-Mouse Lgr5/GPR49 PE-conjugated Monoclonal Antibody (Catalog # FAB8240P) or (B) Rat IgG2B PE-conjugated Isotype Control (Catalog # IC013P). (A) Cells in the upper right

quadrant are double-positive for eGFP and Lgr5/GPR49.

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Human Lgr5/GPR49 Antibody Promotes Canonical Wnt Signaling. Stimulation of Lgr5- expressing HEK293 human embryonic kidney cells with Wnt-3a induced TOPflash Luciferase activity, indicating increased b-Catenin-dependent Wnt signaling. Both Rat Anti-Mouse Lgr5/ GPR49 Monoclonal Antibody (Catalog # MAB8240; published as clone RD20) and Mouse Anti-Human Lgr5/GPR49 Monoclonal Antibody (Catalog # MAB8078; published as clone RD42) enhanced Wnt-3a-induced TOPflash Luciferase activity (Data courtesy of Dr. Wim de Lau and Dr. Hans Clevers, Hubrecht Institute, The Netherlands). See also Peng et al. (2013) Cell Rep. 3(6):1885.

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Lgr5/GPR49 is Localized to the Cell Surface and Cytoplasm. The HEK293 human embryonic kidney cell line was transfected with GFP-tagged mouse Lgr5/GPR49 (green). Cells were immersion fixed and stained with Rat Anti-Mouse Lgr5/GPR49 Monoclonal Antibody (red; Catalog # MAB8240) followed by NorthernLights™ 557-conjugated Anti-Rat

IgG Secondary Antibody (Catalog # NL013). The LGR5/GPR49 antibody and the GFP-tagged Lgr5/GPR49 colocalized (yellow) at the cell surface and within the cytoplasm. The cells were counterstained with DAPI (blue).

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R-Spondin Neutralizing Antibodies: Modulate Wnt Signaling

Human R-Spondin 2 Antibody Neutralizes the b-Catenin Response Induced by Recombinant R-Spondin 2. Recombinant Human (rh)R-Spondin 2 (30 ng/mL; Catalog # 3266RS) potentiates Wnt-3a (5 ng/mL; Catalog #1324-WN) induced b-catenin activation in HEK293T human embryonic kidney cells (orange line). Rat Anti-Human R-Spondin 2 Monoclonal Antibody (Catalog # MAB3266) neutralized this response in a dose-dependent manner.

Antibodies for Wnt Research

ROR1 and ROR2: Now Detectable by Flow Cytometry!

Detection of Human ROR1 by Flow Cytometry. MDA MB 231 human breast cancer cell line was stained with Goat Anti-Human ROR1 Alexa Fluor® 488-conjugated Antigen Affinity-

purified Polyclonal Antibody (filled histogram; Catalog # FAB2000G) or Alexa Fluor 488-conjugated Goat IgG Isotype Control Antibody (open; Catalog # IC108G).

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Detection of Human ROR2 by Flow Cytometry. SH SY5Y human neuroblastoma cell line was stained with Mouse Anti-Human ROR2 PE-conjugated Monoclonal Antibody (filled histogram; Catalog # FAB20641P) or PE-conjugated Mouse IgG2A Isotype Control Antibody (open histogram; Catalog # IC003P).

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Tocris Small Molecules for Wnt Signaling

Wnt-C59 - Catalog # 5148

Wnt-C59 is a highly potent Porcupine (PORCN) inhibitor (IC50 = 74 pM). The compound

downregulates Wnt/b-catenin target genes and blocks progression of mammary tumors in MMTV-WNT1 transgenic mice. Wnt-C59 also induces cardiomyocyte differentiation from human iPSCs following culture with CHIR 99021 (Catalog # 4423). The compound is cell permeable and orally bioavailable.

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endo-IWR 1 - Catalog #3532

endo-IWR 1 is an inhibitor of the Wnt/b-catenin signaling pathway. The compound increases Axin-2 protein levels and promotes b-catenin phosphorylation by stabilizing Axin-scaffolded destruction complexes. Its negative control, exo-IWR (Catalog # 3947), is also available.

Full Wnt Family Product Listing