Recombinant Mouse Wnt-3a, Biotinylated Protein

Carrier Free

Catalog # Availability Size / Price Qty
BT1324-010/CF

With Carrier

Catalog # Availability Size / Price Qty
BT1324-010
Recombinant Mouse Wnt-3a, Biotinylated Protein Binding Activity
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Recombinant Mouse Wnt-3a, Biotinylated Protein Summary

Product Specifications

Purity
>75%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.
Endotoxin Level
<1.0 EU per 1 μg of the protein by the LAL method.
Activity
Measured by its binding ability in a functional ELISA. In a 100 µL reaction mixture containing biotinylated Recombinant Mouse Wnt-3a at 50 ng/mL and Recombinant Human Frizzled‑5 Fc Chimera (Catalog # 1617-FZC) dilutions, the concentration of Recombinant Human Frizzled‑5 Fc Chimera that produces 50% of the maximal binding response is 2-12 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.
Source
Chinese Hamster Ovary cell line, CHO-derived mouse Wnt-3a protein
Ser19-Lys352
Accession #
N-terminal Sequence
Analysis
Ser19
Structure / Form
Biotinylated protein via amines
Predicted Molecular Mass
37 kDa (unlabeled)

Product Datasheets

BT1324 (with carrier)

BT1324/CF (carrier free)

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.

BT1324

Formulation Lyophilized from a 0.2 μm filtered solution in PBS, EDTA and CHAPS with BSA as a carrier protein.
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.

BT1324/CF

Formulation Lyophilized from a 0.2 μm filtered solution in PBS, 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.

Data Image

Binding Activity Recombinant Mouse Wnt-3a, Biotinylated Protein Binding Activity View Larger

Recombinant Biotinylated Mouse Wnt-3a (Catalog # BT1324) binds to Recombinant Human Frizzled-5 Fc Chimera (Catalog # 1617-FZC) with an ED50of 2-12 ng/mL.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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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
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

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