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Human/Mouse Wnt-3a Antibody

Catalog #: MAB1324
12 Citations Datasheet / COA / SDS
Newer Version Available: MAB9025
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Discontinued Product

MAB1324 has been discontinued and is replaced by MAB9025. MAB13242.

Detection of Human and Mouse Wnt‑3a by Western Blot.
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Citations (12)
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Human/Mouse Wnt-3a Antibody Summary

Species Reactivity
Human, Mouse
Specificity
Detects mouse Wnt-3a in direct ELISAs and human and mouse Wnt-3a in Western blots. In direct ELISAs and Western blots, this antibody does not cross-react with recombinant mouse Wnt-1, -4, or 5a.
Source
Monoclonal Rat IgG2A Clone # 217804
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
E. coli-derived recombinant mouse Wnt‑3a
Ser36-Gln75 and Trp219-Arg269
Accession # P27467.1
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
Endotoxin Level
<0.10 EU per 1 μg of the antibody by the LAL method.
Label
Unconjugated

Applications

Recommended Concentration
Sample
Western Blot
2 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/106 cells
See below
Neutralization
Measured by its ability to neutralize Wnt‑3a-induced alkaline phosphatase production in the MC3T3‑E1 mouse preosteoblast cell line. The Neutralization Dose (ND50) is typically 3-15 µg/mL in the presence of 10 ng/mL Recombinant Mouse Wnt‑3a or 20 ng/mL Recombinant Human Wnt‑3a.

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.

Scientific Data

Western Blot Detection of Human and Mouse Wnt-3a antibody by Western Blot. View Larger

Detection of Human and Mouse Wnt‑3a by Western Blot. Western blot shows lysates of BG01V human embryonic stem cells and mouse lung tissue. PVDF membrane was probed with 2 µg/mL of Rat Anti-Human/Mouse Wnt-3a Monoclonal Antibody (Catalog # MAB1324) 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 Wnt-3a antibody in BG01V Human Stem Cells antibody by Flow Cytometry. View Larger

Detection of Wnt-3a in BG01V Human Stem Cells by Flow Cytometry. BG01V human embryonic stem cells were stained with Rat Anti-Human/Mouse Wnt-3a Monoclonal Antibody (Catalog # MAB1324, filled histogram) or isotype control antibody (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 Fixation & Permeabilization Buffer Kit (Catalog # FC012).

Neutralization Alkaline Phosphatase Production Induced by Wnt‑3a and Neutralization by Human/Mouse Wnt‑3a Antibody. View Larger

Alkaline Phosphatase Production Induced by Wnt‑3a and Neutralization by Human/Mouse Wnt‑3a Antibody. Recombinant Mouse Wnt-3a (Catalog # 1324-WN) induces alkaline phosphatase production in the MC3T3-E1 mouse preosteoblast cell line in a dose-dependent manner (orange line). Under these conditions, alkaline phosphatase production elicited by Recombinant Mouse Wnt-3a (10 ng/mL) is neutralized (green line) by increasing concentrations of Rat Anti-Human/Mouse Wnt-3a Monoclonal Antibody (Catalog # MAB1324). The ND50 is typically 3-15 µg/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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Preparation and Storage

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
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Shipping
Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store 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.
  • 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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Citations for Human/Mouse Wnt-3a Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

12 Citations: Showing 1 - 10
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  1. Wnt/&beta-catenin signaling regulates brain-derived neurotrophic factor release from spinal microglia to mediate HIV1 gp120-induced neuropathic pain
    Authors: X Zhou, L Tao, M Zhao, S Wu, E Obeng, D Wang, W Zhang
    Mol Pain, 2020-01-01;16(0):1744806920922.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC
  2. Bone metastasis of prostate cancer can be therapeutically targeted at the TBX2-WNT signaling axis
    Authors: S Nandana, M Tripathi, P Duan, CY Chu, R Mishra, C Liu, R Jin, H Yamashita, M Zayzafoon, NA Bhowmick, HE Zhau, RJ Matusik, LW Chung
    Cancer Res, 2017-01-20;0(0):.
    Species: Human
    Sample Types: Whole Cells
    Applications: Neutralization
  3. SULF2 overexpression positively regulates tumorigenicity of human prostate cancer cells.
    Authors: Vicente C, Lima M, Nader H, Toma L
    J Exp Clin Cancer Res, 2015-03-14;34(0):25.
    Species: Human
    Sample Types: Whole Cells
    Applications: Flow Cytometry
  4. (Pro)renin receptor is crucial for Wnt/beta-catenin-dependent genesis of pancreatic ductal adenocarcinoma.
    Authors: Shibayama Y, Fujimori T, Nguyen G, Hirose T, Totsune K, Ichihara A, Kitada K, Nakano D, Kobori H, Kohno M, Masaki T, Suzuki Y, Yachida S, Nishiyama A
    Sci Rep, 2015-03-09;5(0):8854.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  5. Drugging a stem cell compartment using Wnt3a protein as a therapeutic.
    Authors: Dhamdhere, Girija R, Fang, Mark Y, Jiang, Jie, Lee, Katherin, Cheng, Du, Olveda, Rebecca, Liu, Bo, Mulligan, Kimberle, Carlson, Jeffery, Ransom, Ryan C, Weis, William, Helms, Jill A
    PLoS ONE, 2014-01-06;9(1):e83650.
    Species: Human, Mouse
    Sample Types: Recombinant Protein
    Applications: Western Blot
  6. Influence of in vitro and in vivo oxygen modulation on beta cell differentiation from human embryonic stem cells.
    Authors: Cechin S, Alvarez-Cubela S, Giraldo J, Molano R, Villate S, Ricordi C, Pileggi A, Inverardi L, Fraker C, Dominguez-Bendala J
    Stem Cells Transl Med, 2013-12-27;3(3):277-89.
    Species: Human
    Sample Types: Cell Culture Supernates
    Applications: Bioassay
  7. Secreted Frizzled-related protein potentiation versus inhibition of Wnt3a/beta-catenin signaling.
    Authors: Xavier C, Melikova M, Chuman Y, Uren A, Baljinnyam B, Rubin J
    Cell Signal, 2013-09-28;26(1):94-101.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Co-Immunoprecipitation
  8. Extracellular calcium-sensing receptor/PTH knockout mice colons have increased Wnt/beta-catenin signaling, reduced non-canonical Wnt signaling, and increased susceptibility to azoxymethane-induced aberrant crypt foci.
    Authors: MacLeod R
    Lab Invest, 2013-04-01;93(5):520-7.
    Species: Mouse
    Sample Types: Tissue Homogenates
    Applications: Western Blot
  9. Impaired skeletal muscle regeneration in the absence of fibrosis during hibernation in 13-lined ground squirrels.
    Authors: Andres-Mateos, Eva, Mejias, Rebeca, Soleimani, Arshia, Lin, Brian M, Burks, Tyesha N, Marx, Ruth, Lin, Benjamin, Zellars, Richard, Zhang, Yonggang, Huso, David L, Marr, Tom G, Leinwand, Leslie A, Merriman, Dana K, Cohn, Ronald D
    PLoS ONE, 2012-11-14;7(11):e48884.
    Species: Squirrel
    Sample Types: Tissue Homogenates
    Applications: Western Blot
  10. Cooperative folding and ligand-binding properties of LRP6 beta-propeller domains.
    Authors: Liu CC, Pearson C, Bu G
    J. Biol. Chem., 2009-03-31;284(22):15299-307.
    Species: Mouse
    Sample Types: Cell Lysates
    Applications: Western Blot
  11. Prostate tumor progression is mediated by a paracrine TGF-beta/Wnt3a signaling axis.
    Authors: Li X, Placencio V, Iturregui JM, Uwamariya C, Sharif-Afshar AR, Koyama T, Hayward SW, Bhowmick NA
    Oncogene, 2008-08-25;27(56):7118-30.
    Species: Mouse
    Sample Types: Cell Lysates, In Vivo
    Applications: Neutralization, Western Blot
  12. Nephroblastoma overexpressed (Nov) inhibits osteoblastogenesis and causes osteopenia.
    Authors: Rydziel S, Stadmeyer L, Zanotti S, Durant D, Smerdel-Ramoya A, Canalis E
    J. Biol. Chem., 2007-05-10;282(27):19762-72.
    Species: Mouse
    Sample Types: Protein
    Applications: Western Blot

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