Can I reconstitute Recombinant Human Wnt-3a (High Purity) Protein (catalog # 5036-WNP) at a different concentration than recommended?

Name: Recombinant Human Wnt-3a (High Purity) Protein
Brand: R&D Systems
SKU: 5036-WNP

We cannot guarantee the performance if the protein is reconstituted at a different concentration than recommended on the product datasheet. Please contact Technical Service if you have any questions or concerns about reconstitution.

Recombinant Human Wnt-3a (High Purity) Protein

R&D Systems, Inc. a Bio-Techne Brand

Catalog # 5036-WNP

Citations (4)

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Key Product Details

Source:	CHO
Accession #:	NP_149122
Applications:	Bioactivity

View Full Technical Details for Recombinant Human Wnt-3a (High Purity) Protein

Product Specifications for Recombinant Human Wnt-3a (High Purity) Protein

Source	Chinese Hamster Ovary cell line, CHO-derived human Wnt-3a protein Ser19-Lys352
Purity	>90%, by SDS-PAGE with silver staining
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. The ED₅₀ for this effect is 1.5-7.5 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.

Scientific Data Examples for Recombinant Human Wnt-3a (High Purity) Protein

	Recombinant Human Wnt-3a (High Purity) Protein Bioactivity Recombinant Human Wnt-3a (High Purity, Catalog # 5036-WNP) induces alkaline phosphatase production by the MC3T3-E1 mouse preosteoblast cell line. The ED₅₀ for this effect is 1.5-7.5 ng/mL.
	Recombinant Human Wnt-3a (High Purity) Protein SDS-PAGE 1 μg/lane of Recombinant Human Wnt-3a (High Purity) was resolved with SDS-PAGE under reducing (R) conditions and visualized by silver staining, showing a single band at 40 kDa.

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.

5036-WNP

Carrier

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

5036-WNP/CF

Carrier Free

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

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). 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).

Item	Value
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, Wnt-3a, Wnt3a, protein Wnt-3a, wingless-type MMTV integration site family, member 3A

Citations for Recombinant Human Wnt-3a (High Purity) Protein (4)

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Citations are publications that use Bio-Techne products. Selected citations for Recombinant Human Wnt-3a (High Purity) Protein include:

Species: Human
Sample Types: Whole Cells
Applications: Bioassay

J Krumm et al. (2022), High temporal resolution proteome and phosphoproteome profiling of stem cell-derived hepatocyte development Cell Reports, 38(13):110604.
PMID: 35354033
Species: Human
Sample Types: Whole Cells
Applications: Bioassay

Bilir B et al. (2013), Wnt signaling blockage inhibits cell proliferation and migration, and induces apoptosis in triple-negative breast cancer cells. J Transl Med, 11(0):280.
PMID: 24188694
Species: Human
Sample Types: Recombinant Protein
Applications: Binding Assay

Bourhis E et al. (2010), Reconstitution of a frizzled8.Wnt3a.LRP6 signaling complex reveals multiple Wnt and Dkk1 binding sites on LRP6. J. Biol. Chem., 285(12):9172-9.
PMID: 20093360
Species: Mouse
Sample Types: Whole Cells
Applications: Bioassay

Carthy J et al. (0), Commercially Available Preparations of Recombinant Wnt3a Contain Non-Wnt Related Activities Which May Activate TGF-beta Signaling. J Cell Bioche117(4):938-45.
PMID: 26369756

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Showing 1 - 4 of 4 citations Showing All

FAQs for Recombinant Human Wnt-3a (High Purity) Protein

Can I reconstitute Recombinant Human Wnt-3a (High Purity) Protein (catalog # 5036-WNP) at a different concentration than recommended?

We cannot guarantee the performance if the protein is reconstituted at a different concentration than recommended on the product datasheet. Please contact Technical Service if you have any questions or concerns about reconstitution.

Recombinant Human Wnt-3a (High Purity) Protein

R&D Systems, Inc. a Bio-Techne Brand

Key Product Details

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Product Specifications for Recombinant Human Wnt-3a (High Purity) Protein

Scientific Data Examples for Recombinant Human Wnt-3a (High Purity) Protein

Formulation, Preparation and Storage

Carrier Free

5036-WNP

Carrier

5036-WNP/CF

Carrier Free

Reconstitution Calculator

Background: Wnt-3a

Citations for Recombinant Human Wnt-3a (High Purity) Protein (4)

FAQs for Recombinant Human Wnt-3a (High Purity) Protein

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

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