Recombinant Mouse Wnt-3a (High Purity) Protein

R&D Systems, part of Bio-Techne | Catalog # 1324-WNP

R&D Systems, part of Bio-Techne
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Carrier Free
1324-WNP-010/CF
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With Carrier
1324-WNP-010
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Key Product Details

Source

CHO

Accession #

P27467

Conjugate

Unconjugated

Applications

Bioactivity

View all Wnt-3a Proteins and Enzymes »

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived mouse Wnt-3a protein
Ser19-Lys352

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

<0.10 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. Fischer, L. et al. (2002) J. Biol. Chem. 277:30870.
The ED50 for this effect is ≤5 ng/mL.

Measured by its ability to induce Topflash reporter activity in HEK293T human embryonic kidney cells.

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.

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: 1324-WNP
Formulation Lyophilized from a 0.2 μm filtered solution in PBS, NaCl, EDTA and CHAPS with BSA as a carrier protein.
Reconstitution Reconstitute at 100 μg/mL in 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.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.
Carrier Free: 1324-WNP/CF
Formulation Lyophilized from a 0.2 μm filtered solution in PBS, NaCl, EDTA and CHAPS.
Reconstitution Reconstitute at 100 μg/mL in 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.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, -20 to -70 °C under sterile conditions after reconstitution.

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). Mouse Wnt-3a shares 96% amino acid (aa) identity with human Wnt-3a, and 97% with bovine and canine Wnt-3a. The rat Wnt-3a precursor shares 100% aa identity with mouse Wnt-3a aa 63-352 (8), and 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 (13-15).

References

  1. Willert, K. and Nusse, R. (2012) Cold Spring Harb. Perspect. Biol. 4:a007864.
  2. MacDonald, B.T. and X. He (2012) Cold Spring Harb. Perspect. Biol. 4:a007880.
  3. Korinek, V. et al. (1997) Science 275:1784.
  4. Chen, B. et al. (2009) Nat. Chem. Biol. 5:100.
  5. Smolich, B.D. et al. (1993) Mol. Biol. Cell 4:1267.
  6. Takada, R. et al. (2006) Dev. Cell 11:791.
  7. Komekado, H. (2007) Genes Cells 12:521.
  8. Entrez Accession # NP_001100475.
  9. Dunty Jr. W. C. et al. (2008) Development 135:85.
  10. Ikeya, M. and S. Takada (2001) Mech. Dev. 103:27.
  11. Lee, S. M. et al. (2000) Development 127:457.
  12. Takada, S. et al. (1994) Genes Dev. 8:174.
  13. Willert, K. et al. (2003) Nature 423:6938.
  14. Kalani, M.Y. et al. (2008) Proc. Natl. Acad. Sci. USA 105:16970.
  15. Ten Berge, D. et al. (2011) Nat. Cell Biol. 13:1070.

Long Name

Wingless-type MMTV Integration Site Family, Member 3a

Alternate Names

Wnt3a

Entrez Gene IDs

89780 (Human); 22416 (Mouse)

Gene Symbol

WNT3A

UniProt

P27467

Additional Wnt-3a Products

Product Documents for Recombinant Mouse Wnt-3a (High Purity) Protein

Product Datasheets

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Download SDS 1324-WNP (with carrier)
Download SDS 1324-WNP/CF (carrier free)

Product Specific Notices for Recombinant Mouse Wnt-3a (High Purity) Protein

For research use only

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