Recombinant Human Wnt-3a GMP Protein, CF

Catalog #: 5036-GMP
1 Citation Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
5036-GMP-010
5036-GMP-01M
5036-GMP-050

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Recombinant Human Wnt-3a GMP Protein Bioactivity

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Product Details

Citations (1)

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human Wnt-3a GMP Protein, CF Summary

Product Specifications

Purity

>75%, by SDS-PAGE with silver staining, under reducing conditions

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

Activity

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.

Source

Chinese Hamster Ovary cell line, CHO-derived human Wnt-3a protein
Ser19-Lys352
Manufactured and tested under cGMP guidelines.

Accession #

NP_149122.1

N-terminal Sequence
Analysis

Ser₁₉-Tyr-Pro-Ile-Trp-Trp-Ser-Leu-Ala-Val

Predicted Molecular Mass

37.4 kDa

SDS-PAGE

40 kDa, reducing conditions

Product Datasheets

5036-GMP

Product Datasheet

COA

SDS

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.

5036-GMP

Formulation	Lyophilized from a 0.2 μm filtered solution in PBS, EDTA and CHAPS.
Reconstitution	Reconstitute at 200 μg/mL in PBS.
Shipping	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 12 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.

Scientific Data

Bioactivity

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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.

Bioactivity

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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.

Reconstitution Calculator

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

Entrez Gene IDs

89780 (Human); 22416 (Mouse)

Alternate Names

MGC119418; MGC119419; MGC119420; protein Wnt-3a; wingless-type MMTV integration site family, member 3A; Wnt3a; Wnt-3a

Manufacturing Specifications

GMP Proteins
R&D Systems, a Bio-Techne Brand's GMP proteins are produced according to relevant sections of the following documents: USP Chapter 1043, Ancillary Materials for Cell, Gene and Tissue-Engineered Products and Eu. Ph. 5.2.12, Raw Materials of Biological Origin for the Production of Cell-based and Gene Therapy Medicinal Products.

R&D Systems' quality focus includes:

Designed, manufactured and tested under an ISO 9001:2015 and ISO 13485:2016 certified quality system
Documented and controlled manufacturing process
Control of documentation and process changes by QA
Personnel training programs
Raw material inspection and vendor qualification/monitoring program
Validated equipment, processes and test methods
Equipment calibration and maintenance schedules using a Regulatory Asset Manager
Facility/Utilities maintenance, contamination controls, safety and pest control programs
Material review process for variances
Robust product stability program following relevant ICH guidelines

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 application. Each product is provided with a lot-specific Certificate of Analysis that contains the product's specifications and test results. Quality control testing may include, but is not limited to:

N-terminal amino acid analysis
SDS-PAGE purity analysis
Molecular weight analysis via mass spectrometry
Endotoxin assessment per USP <85> and Ph. Eur. 2.6.14 guidelines
Bioassay analysis
Microbial testing per USP <71> and Ph. Eur. 2.6.1 guidelines
Host cell protein assessment
Host cell DNA assessment
Mycoplasma assessment

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 and St. Paul, 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

Full terms and conditions of sale can be found online in the Protein Sciences Segment T&Cs at: Terms & Conditions.

Related Research Areas

Citation for Recombinant Human Wnt-3a GMP Protein, CF

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1