Wnt-3a Antibody (217804.2R) [Unconjugated]

Western blot shows lysates of CHO Chinese hamster ovary cell line either mock transfected or transfected with mouse Wnt-3a. PVDF membrane was probed with 1 µg/mL of Rat Anti-Human/Mouse Wnt‑3a Monoclonal Antibody (Catalog # MAB13242) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). A specific band was detected for Wnt‑3a at approximately 40 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

BGO1v human embryonic stem cells were stained with Rat Anti-Human/Mouse Wnt-3a Monoclonal Antibody (Catalog # MAB13242, filled histogram) or isotype controlantibody (Catalog # MAB006, open histogram), followed by Allophycocyanin-conjugated Anti-Rat IgG Secondary Antibody (Catalog # F0113). To facilitate intracellular staining, cells were fixed and permeabilized with FlowX FoxP3/Transcription Factor Fixation & Permeabilization Buffer Kit (Catalog # FC012).

• Reactivity: Hu, Mu
• Applications: WB, CyTOF-ready, Flow
• Clone: 217804.2R
• Clonality: Monoclonal
• Host: Rat
• Conjugate: Unconjugated

Wnt-3a Antibody (217804.2R) [Unconjugated] Summary

• Additional Information: Recombinant Monoclonal Antibody.
• Immunogen: E.coli-derived recombinant mouse Wnt-3a
  Ser36-Gln75, Trp219-Arg269
  Accession # P27467
• Specificity: Detects mouse Wnt-3a in direct ELISAs.
• Details of Functionality: ActivityAssaywCitation not found None None
• Source: N/A
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Rat

Applications/Dilutions

• Dilutions:
  • Western Blot 1 ug/mL
  • CyTOF-ready
  • Intracellular Staining by Flow Cytometry 0.25 ug/mL

Packaging, Storage & Formulations

• 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.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Wnt-3a Antibody (217804.2R) [Unconjugated]

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

Background

Wnt-3a is one of about 19 vertebrate members of the Wingless-type MMTV integration site (Wnt) family of highly conserved cysteine-rich secreted glycoproteins important for normal developmental processes (1‑3). Wnts bind to receptors of the Frizzled family in conjunction with a coreceptor of the low-density lipoprotein receptor-related protein family (LRP-5 or -6), or the Ryk atypical receptor tyrosine kinase (1, 4). Mouse Wnt-3a is a 44 kDa secreted hydrophobic glycoprotein containing a conserved pattern of 24 cysteine residues (5). Like other Wnts, Wnt-3a is modified by palmitate addition (at Cys 77) following glycosylation, which increases its hydrophobicity, secretion and activity (6, 7). A second site at Ser 209 is modified by palmitoleic acid and also contributes to activity and secretion (8). 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 as it is apparently translated shares 100% aa identity with mouse Wnt-3a aa 63-352 (9). Wnt-3a also shares 87% aa identity with Wnt-3. During development, Wnt-3a is morphogen that is thought to coordinate somitogenesis and mesoderm boundary determination, and is expressed at the same locations and times as Wnt-2b and Wnt-5a (10). When Wnt-3a is deleted, mice fail to develop a hippocampus, and show defects in anterior-posterior patterning, somite development and tailbud formation (10-13). Recombinant Wnt-3a promotes proliferation of committed stem cell lineages in vitro, and may help maintain the cells in an undifferentiated state (6, 14) For example, Wnt-3a can induce self-renewal of hematopoietic stem cells, allowing expansion without further differentiation (6).

1. Mikels, A.J. and R. Nusse (2006) Oncogene 25:7461.
2. Miller, J.R. (2001) Genome Biol. 3:3001.
3. Roelink, H. and R. Nusse (1991) Genes Dev. 5:381.
4. Lu, W et al. (2004) Cell 119:97.
5. Smolich, B.D. et al. (1993) Mol. Biol Cell 4:1267.
6. Willert, K. et al. (2003) Nature 423:448.
7. Komekado, H. et al. (2007) Genes Cells 12:521.
8. Takada, R. et al. (2006) Dev. Cell 11:791.
9. Entrez Accession # NP_001100475.
10. Dunty, W.C. et al. (2008) Development 135:85.
11. Lee, S.M.K. et al. (2000) Development 127:457.
12. Takada, S. et al. (1994) Genes Dev. 8:174.
13. Ikeya, M. and S. Takada (2001) Mech. Dev. 103:27.
14. Nusse, R. (2008) Cell Res. 18:523.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Bioinformatics

• Uniprot:
  • P27467()