Human/Mouse Wnt-3a Antibody

Recombinant Monoclonal Antibody
Catalog # Availability Size / Price Qty
MAB13242-100
MAB13242-SP
Detection  of Mouse Wnt‑3a by Western Blot.
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Human/Mouse Wnt-3a Antibody Summary

Species Reactivity
Human, Mouse
Specificity
Detects mouse Wnt-3a in direct ELISAs.
Source
Recombinant Monoclonal Rat IgG2A Clone # 217804.2R
Purification
Protein A or G purified from cell culture supernatant
Immunogen
E.coli-derived recombinant mouse Wnt-3a
Ser36-Gln75, Trp219-Arg269
Accession # P27467
Formulation
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.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 
Intracellular Staining by Flow Cytometry
0.25 µg/mL
See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Data Examples

Western Blot Detection  of Mouse Wnt‑3a by Western Blot. View Larger

Detection of Mouse Wnt‑3a by Western Blot. 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.

Intracellular Staining by Flow Cytometry Detection  of Wnt3A in BG01V Human Stem Cells by Flow Cytometry. View Larger

Detection of Wnt3A in BG01V Human Stem Cells by Flow Cytometry. 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).

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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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
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.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Wnt-3a

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).

References
  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.
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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