Chinese Hamster Ovary cell line, CHO-derived human Wnt-3a protein Ser19-Lys352 Manufactured and tested under cGMP guidelines.
>75%, by SDS-PAGE with silver staining, under reducing conditions.
<0.10 EU per 1 μg of the protein by the LAL method.
N-terminal sequence Analysis
Predicted Molecular Mass
40 kDa, reducing conditions
Measured by its ability to induce alkaline phosphatase production by MC3T3‑E1 mouse preosteoblast cells. The ED50 for this effect is 5-25 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.
Recombinant Human Wnt-3a GMP Protein, CF Scientific Data Examples
Recombinant Human Wnt-3a GMP Protein Bioactivity
GMP-grade Recombinant Human Wnt-3a (Catalog # 5036-GMP) induces alkaline phosphatase production by the MC3T3‑E1 mouse preosteoblast cell line. The ED50 for this effect is 5-25 ng/mL. Three independent lots were tested for activity and plotted on the same graph to show lot-to-lot consistency of GMP Wnt-3a.
Recombinant Human Wnt-3a GMP Protein Bioactivity
GMP-grade Recombinant Human Wnt-3a (Catalog # 5036-GMP) induces beta -catenin-responsive transcriptional activation in a Topflash Luciferase assay using the HEK293T human embryonic kidney cell line. The ED50 for this effect is <500 ng/mL. Three independent lots were tested for activity and plotted on the same graph to show lot-to-lot consistency of GMP Wnt-3a.
Formulation, Preparation and Storage
What does CF mean?
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our
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
Lyophilized from a 0.2 μm filtered solution in PBS, EDTA and CHAPS.
Reconstitute at 200 μg/mL in PBS.
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
A minimum of 6 months when stored at ≤ -20 °C as supplied. Refer to lot specific COA for the Use by Date.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
3 months, ≤ -20 °C under sterile conditions after reconstitution.
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).
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.
Wingless-type MMTV Integration Site Family, Member 3a
Entrez Gene IDs
89780 (Human); 22416 (Mouse)
MGC119418, MGC119419, MGC119420, Wnt-3a, Wnt3a, protein Wnt-3a, wingless-type MMTV integration site family, member 3A
GMP Proteins R&D Systems, a Bio-Techne Brand's GMP proteins are produced according to relevant sections of the following documents: WHO TRS, No. 822, 1992 Annex 1, Good Manufacturing Practices for Biological Products; USP Chapter 1043, Ancillary Materials for Cell, Gene and Tissue-Engineered Products and USP Chapter 92, Growth Factors and Cytokines Used in Cell Therapy Manufacturing.
R&D Systems' quality focus includes:
Manufactured and tested under an ISO 9001:2015 and ISO 13485:2016 certified quality system
Documented processes and QA control of documentation and process changes
Personnel training programs
Raw material testing and vendor qualification/monitoring
Fully validated equipment, processes and test methods
Equipment calibration schedules using a computerized calibration program
Facility maintenance, safety programs and pest control
Material review process for variances
Monitoring of stability over product shelf-life
R&D Systems strives to provide our customers with the analytical characteristics of each product so that customers may determine whether our products are appropriate for their research. The Certificate of Analysis provided contains the following lot specific information:
N-terminal amino acid analysis, SDS-PAGE analysis, and endotoxin level (as determined by LAL assay) performed on each bulk QC lot, not on individual bottlings of each QC lot
Post-bottling lot-specific bioassay results (compliance with an established range) and results of microbial testing according to USP
Host Cell Protein testing performed by ELISA
Mycoplasma testing by ribosomal RNA hybridization assay
Additional testing and documentation requested by the customer can be arranged at an additional cost.
Production records and facilities are available for examination by appropriate personnel on-site at R&D Systems in Minneapolis, Minnesota USA.
R&D Systems sells GMP grade products for preclinical or clinical ex vivo use. They are not for in vivo use. Please read the following End User Terms prior to using this product.
Product Specific Notices
The End User is aware that R&D Systems, Inc. sells GMP products for preclinical or clinical ex vivo use and not for in vivo use. The End User further agrees, as a condition of the sale of R&D Systems' GMP products that: a) the End User will not use this GMP Product in any procedure wherein the product may be directly or indirectly administered to humans, unless the End User has obtained, or prior to their use will have obtained, an Investigational New Drug (IND) exemption from the FDA and will use the product only in accordance with the protocols of such IND and of the Institutional Review Board overseeing the proposed research, or b) the End User will use the products outside of the United States in accordance with the protocols of research approved by the Institutional Review Board or authorized ethics committee and regulatory agencies to which the End User is subject to in their territory.
TERMS AND CONDITIONS
The following limitation applies to R&D Systems' warranty and liability for damages: All products are warranted to meet R&D Systems' published specifications when used under normal laboratory conditions.
R&D SYSTEMS DOES NOT MAKE ANY OTHER WARRANTY OR REPRESENTATION WHATSOEVER, WHETHER EXPRESS OR IMPLIED, WITH RESPECT TO ITS PRODUCTS. IN PARTICULAR, R&D SYSTEMS DOES NOT MAKE ANY WARRANTY OF SUITABILITY, NONINFRINGEMENT, MERCHANTABILITY OR FITNESS FOR ANY PARTICULAR PURPOSE.
NOTWITHSTANDING ANY OTHER PROVISIONS OF THESE TERMS AND/OR ANY OTHER AGREEMENT BETWEEN R&D SYSTEMS AND PURCHASER FOR THE PURCHASE OF THE PRODUCTS, R&D SYSTEMS' TOTAL LIABILITY TO PURCHASER ARISING FROM OR IN RELATION TO THESE TERMS, AN AGREEMENT BETWEEN THE PARTIES OR THE PRODUCTS, WHETHER ARISING IN CONTRACT, TORT OR OTHERWISE SHALL BE LIMITED TO THE TOTAL AMOUNT PAID BY PURCHASER TO R&D SYSTEMS FOR THE APPLICABLE PRODUCTS. IN NO EVENT WILL R&D SYSTEMS BE LIABLE FOR THE COST OF PROCUREMENT OF SUBSTITUTE GOODS.
Which Brands are Currently Available on bio‑techne.com? R&D Systems, Tocris Bioscience and ProteinSimple branded products are available to purchase through bio‑techne.com. ProteinSimple branded instruments are available to quote. ACD branded products will be available on bio‑techne.com in the near future. Novus Biologicals branded products are not currently available on bio‑techne.com and can be found at novusbio.com.
Setting the standard in quality research reagents for over 30 years
A trusted leader in quality life science reagents
Your trusted supplier for innovative and high performance life science reagents
Proprietary systems and consumables for simpler, more quantitative and affordable protein analysis
Proprietary RNAscope® technology capable of detecting and quantifying RNA biomarkers in situ at single molecule sensitivity
A world leader in developing liquid biopsy based diagnostics