Recombinant Human Wnt-3a Protein Best Seller
R&D Systems, part of Bio-Techne | Catalog # 5036-WN
Key Product Details
Product Specifications
Source
Chinese Hamster Ovary cell line, CHO-derived human Wnt-3a protein
Ser19-Lys352
Ser19-Lys352
Purity
>75%, by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
N-terminal Sequence Analysis
Ser19
Predicted Molecular Mass
37.4 kDa
SDS-PAGE
40 kDa, reducing conditions
Activity
Measured by its ability to induce alkaline phosphatase production by MC3T3-E1 mouse preosteoblast cells.
The ED50 for this effect is 5.00-25.0 ng/mL.
Measured by its ability to induce Topflash reporter activity in HEK293T human embryonic kidney cells.
The ED50 for this effect is <500 ng/mL. Protein concentrations should be titrated based on cell type and if appropriate, passage number of the cell line.
Optimal concentrations should be determined by each laboratory for each application.
The ED50 for this effect is 5.00-25.0 ng/mL.
Measured by its ability to induce Topflash reporter activity in HEK293T human embryonic kidney cells.
The ED50 for this effect is <500 ng/mL. Protein concentrations should be titrated based on cell type and if appropriate, passage number of the cell line.
Optimal concentrations should be determined by each laboratory for each application.
Reviewed Applications
Read 27 reviews rated 4.7 using 5036-WN in the following applications:
- Cell Culture (1 Review)
- Cell migration/motility (2 Reviews)
- Cell Proliferation (5 Reviews)
- Enzymatic activity in vitro (2 Reviews)
- In vitro bioactivity in cell culture (6 Reviews)
- Luciferase reporter activity (1 Review)
- Media additive for protein or antibody production (1 Review)
- SDS-PAGE Control (2 Reviews)
- Stem/Immune cell maintenance or differentiation (7 Reviews)
Scientific Data Images for Recombinant Human Wnt-3a Protein
Recombinant Human Wnt-3a Protein Bioactivity
Recombinant Human Wnt-3a (Catalog # 5036-WN) induces alkaline phosphatase production by the MC3T3-E1 mouse preosteoblast cell line. The ED50 is 1.7-fold better with more than twice the maximum response compared to the top competitor's Wnt-3a.
Recombinant Human Wnt-3a Protein Lot to Lot Consistency
The lot-to-lot consistency of Recombinant Human Wnt-3a Protein (Catalog # 5036-WN) was assessed by testing the ability of three independent lots of the protein to induce alkaline phosphatase production in the MC3T3-E1 mouse preosteoblast cell line. Each trace on the graph represents data obtained from Recombinant Human Wnt-3a from a different manufacturing run. The ED50 for this effect is 5.00-25.0 ng/mL.
iPSC-derived Human Intestinal Organoids Cultured using Recombinant Human Wnt-3a Protein.
iPSC-derived human intestinal organoids were cultured using Cultrex™ UltiMatrix RGF Basement Membrane Extract (BME001-05) and intestinal organoid culture medium, which includes Recombinant Human Wnt-3a (Catalog # 5036-WN), Recombinant Human EGF (236-EG), Recombinant Human Noggin (6057-NG), and Recombinant Human R-Spondin 1 (4645-RS), along with the other reagents listed in the intestinal organoid culture medium recipe in the human intestinal organoid culture protocol. (A) Human intestinal organoids were stained using a Rat Anti-Human/Mouse/Rat Vimentin Monoclonal Antibody (MAB2105; green) and a Goat Anti-Human/Mouse Desmin Antigen Affinity-purified Polyclonal Antibody (AF3844; red) to visualize myofibroblast cells and counterstained with DAPI (5748; blue). (B) Human intestinal organoids were stained using a Goat Anti-Human/Mouse E-Cadherin Antigen Affinity-purified Polyclonal Antibody (AF748; green) and a Mouse Anti-Human MUC2 Monoclonal Antibody (Novus Biologicals, Catalog # NBP2-44431; red) and counterstained with DAPI (5748; blue).Formulation, Preparation and Storage
Carrier Free
What does CF mean?CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
What formulation is right for me?In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
Carrier: 5036-WN
| Formulation | Lyophilized from a 0.2 μm filtered solution in PBS, EDTA and CHAPS with BSA as a carrier protein. |
| Reconstitution | Reconstitute at 200 μg/mL in sterile PBS containing at least 0.1% human or bovine serum albumin. |
| Shipping | The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Carrier Free: 5036-WN/CF
| Formulation | Lyophilized from a 0.2 μm filtered solution in PBS, EDTA and CHAPS. |
| Reconstitution | Reconstitute at 200 μg/mL in sterile PBS. |
| Shipping | The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Background: Wnt-3a
Wnt-3a is one of 19 vertebrate members of the Wingless-type MMTV integration site (Wnt) family of highly conserved cysteine-rich secreted glycoproteins important for normal developmental processes (1). Wnts bind to the cell surface Frizzled family receptors in conjunction with low-density lipoprotein receptor-related protein family receptors (LRP5 or 6) resulting in the stabilization of intracellular beta-catenin levels (2). As intracellular beta-catenin levels rise, beta-catenin binds to TCF/LEF transcription factors leading to expression of Wnt target genes (3). Endo-IWR 1 (Catalog # 3532, # PSM1324) is a cell-permeant small molecule inhibitor of Axin turnover that suppresses Wnt signal transduction by stabilizing the beta-catenin destruction complex (4). Wnt-3a is a 44 kDa secreted hydrophobic glycoprotein containing a conserved pattern of 24 cysteine residues (5). Wnt-3a has two N-linked glycosylation sites (Asn 87, Asn 298), and Ser 209 is modified with palmitoleic acid (6). Glycosylation and acylation are essential for efficient Wnt secretion and biological activity, respectively (6, 7). Human Wnt-3a shares 96% amino acid (aa) identity with mouse mouse, bovine and canine Wnt-3a, and 89%, 86% and 84% aa identity with chicken, Xenopus and zebrafish Wnt-3a, respectively. It also shares 87% aa identity with Wnt3. During embryonic development, Wnt-3a is necessary for proper development of the hippocampus, anterior-posterior patterning, somite development, and tailbud formation (9-12). Wnt-3a also promotes self-renewal of hematopoietic stem cells, neural stem cells, and embryonic stem cells (13, 14).
References
- Willert, K. and Nusse, R. (2012) Cold Spring Harb. Perspect. Biol. 4:a007864.
- MacDonald, B.T. and X. He (2012) Cold Spring Harb. Perspect. Biol. 4:a007880.
- Korinek, V. et al. (1997) Science 275:1784.
- Chen, B. et al. (2009) Nat. Chem. Biol. 5:100.
- Smolich, B.D. et al. (1993) Mol. Biol. Cell 4:1267.
- Takada, R. et al. (2006) Dev. Cell 11:791.
- Komekado, H. (2007) Genes Cells 12:521.
- Dunty Jr. W. C. et al. (2008) Development 135:85.
- Ikeya, M. and S. Takada (2001) Mech. Dev. 103:27.
- Lee, S. M. et al. (2000) Development 127:457.
- Takada, S. et al. (1994) Genes Dev. 8:174.
- Willert, K. et al. (2003) Nature 423:6938.
- Kalani, M.Y. et al. (2008) Proc. Natl. Acad. Sci. USA 105:16970.
- Ten Berge, D. et al. (2011) Nat. Cell Biol. 13:1070.
Long Name
Wingless-type MMTV Integration Site Family, Member 3a
Alternate Names
Wnt3a
Gene Symbol
WNT3A
UniProt
Additional Wnt-3a Products
Product Documents for Recombinant Human Wnt-3a Protein
Product Specific Notices for Recombinant Human Wnt-3a Protein
For research use only
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