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Human/Mouse Wnt-5a Antibody

R&D Systems, part of Bio-Techne | Catalog # MAB645

R&D Systems, part of Bio-Techne
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MAB645-SP
MAB645

Key Product Details

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse, Rat, Xenograft

Applications

Validated:

Immunohistochemistry

Cited:

Bioassay, IHC, IHC-Fr, IHC-P, Neutralization, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Rat IgG2A Clone # 442625

Product Summary for Human/Mouse Wnt-5a Antibody

Immunogen

CHO-derived recombinant mouse Wnt-5a
Gln38-Lys380
Accession # P22725

Specificity

Detects mouse Wnt-5a in direct ELISAs. In direct ELISAs, no cross-reactivity with recombinant mouse Wnt-1, 2b, 3a, 4, 5b, 8a, 8b, 9b, 10a, 10b, 11, or 16 is observed.

Clonality

Monoclonal

Host

Rat

Isotype

IgG2A

Scientific Data Images for Human/Mouse Wnt-5a Antibody

Wnt-5a antibody in Mouse Embryo by Immunohistochemistry (IHC-Fr).

Wnt‑5a in Mouse Embryo.

Wnt-5a was detected in immersion fixed frozen sections of mouse embryo using Human/Mouse Wnt-5a Monoclonal Antibody (Catalog # MAB645) at 10 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Rat IgG Secondary Antibody (orange; Catalog # NL013) and counter-stained with DAPI (blue). View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.
Detection of Mouse Wnt-5a by Immunocytochemistry/Immunofluorescence

Detection of Mouse Wnt-5a by Immunocytochemistry/Immunofluorescence

WNT-5A expression in mouse GFAP+astrocytes. (A) Immunohistochemistry was performed on adult mouse brain sections using an anti-WNT-5A antibody in combination with anti-glial fibrillary acidic protein (GFAP) and ionized calcium-binding adaptor protein1 (IBA1) antibodies as astrocyte and microglia marker, respectively. Merge presents the overlay of IBA1, GFAP, WNT-5A. Size bar −2 μm. The images represent a maximum intensity projection of a Z-stack of 5 μm thickness. The white square marked ‘B’ indicates the area magnified in B: (B) Close up of a GFAP+ astrocyte reveals the expression of WNT-5A in this cell type. (C) shows immunoblot detection of recombinant WNT-5A (rWNT-5A; 375 ng/lane) in comparison to lysates from mouse primary microglia and mixed astrocyte cultures. beta-actin serves as a loading control. (D) The bar graph depicts expression levels of WNT-5A mRNA in mouse primary microglia and mixed astrocyte cultures measured by QPCR. Data are normalized to GAPDH expression and analyzed with a non-parametric Mann–Whitney test. *, P < 0.05; **, P < 0.01; ***, P < 0.001. n = 4 to 8. (E) shows indirect immunocytochemistry of mixed astrocyte cultures employing anti-GFAP as astrocyte and anti-CD11b as microglia markers. DAPI is used as nuclear counterstain. Image represents a maximum image projection of an 8 μm Z-stack. Size bar 20 μm. The frame shows 63 cells in total and 10 CD11b-positive microglia (arrows). Routinely 10% to 18% microglia were observed (n = 4). DAPI, 4',6-diamidino-2-phenylindole; GADPH, glyceraldehyde 3-phosphate dehydrogenase; n, number. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/22647544), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse Wnt-5a by Western Blot

Detection of Mouse Wnt-5a by Western Blot

Wnt5a expressing tumors have less Wnt/ beta-catenin signaling than MMTV-Wnt1 tumors.(A) Quantitative RT-PCR of Wnt/ beta-catenin target genes. Expression of Axin2 mRNA in MMTV-Wnt1 versus MMTV-Wnt1;MMTV-Wnt5a tumors as determined by quantitative RT-PCR (n = 5 MMTV-Wnt1, n = 5 MMTV-Wnt1;MMTV-Wnt5a). Data are shown as tables obtained using REST software. Axin2 mRNA was significantly down-regulated in MMTV-Wnt1;MMTV-Wnt5a tumors. (B) Western blot for beta-catenin protein. Protein lysates were prepared from MMTV-Wnt1 and MMTV-Wnt1;MMTV-Wnt5a tumors. beta-catenin and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were used as loading controls. The ratio of active beta-catenin to beta-catenin as determined by densitometic analysis is shown. MMTV-Wnt1;MMTV-Wnt5a tumors displayed decreased levels of active beta-catenin compared to controls. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0113247), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human/Mouse Wnt-5a Antibody

Application
Recommended Usage

Immunohistochemistry

8-25 µg/mL
Sample: Immersion-fixed frozen sections of mouse embryo
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 2 reviews rated 4 using MAB645 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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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.

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-5a

Wnt proteins are secreted glycoproteins that contain a conserved pattern of 23‑24 cysteine residues. Wnts play critical roles in both carcinogenesis and embryonic development for a variety of organisms. Wnts bind to receptors of the Frizzled family, sometimes in conjunction with other membrane-associated proteins such as LRPs or proteoglycans. Downstream effects of Wnt signaling occur through different intracellular components, depending on which pathway is activated. Three pathways have been characterized: the canonical Wnt/ beta-catenin pathway, the Wnt/Ca2+ pathway, and the planar cell polarity (1‑2).

Wnt-5a is part of the subgroup of Wnts that are not axis-inducing in Xenopus embryos and do not transform C57MG mammary epithelial cells. This subgroup is also implicated in the Wnt/Ca2+ pathway, playing roles in cell movements and cell adhesion (3). This non-canonical Wnt pathway can inhibit canonical Wnt/ beta-catenin signaling. In Wnt-5a deficient mouse embryos, beta-catenin accumulates in the limb bud suggesting that Wnt-5a normally promotes degradation of beta-catenin (4). Likewise, in Xenopus embryos Wnt-5a antagonizes the ability of the canonical Wnt subgroup to induce a secondary axis (5). Wnt-5a is implicated in various types of cancer and has complex roles. It acts as a tumor suppressor for mammary, B-cell, colon, and uroepithelial cancer cells but is up-regulated in melanomas, where expression levels correlate with severity of metastasis (3). Furthermore, aberrant Wnt-5a signaling results in other diseases such as rheumatoid arthritis (6). Like other developmental growth factors Wnt-5a has diverse roles in development. They are too numerous to enunciate here, as functions span from early anterior-posterior development and gastrulation movements to maintaining hematopoietic stem cell population, lung morphogenesis, and limb outgrowth. Mature Wnt-5a is a 49 kDa protein that shares 99% amino acid identity in mouse, rat and human.

References

  1. Miller, J.R. (2002) Genome Biol. 3:3001.
  2. Roelink, H. and R. Nusse (1991) Genes Dev. 5:381.
  3. Veeman, M.T. et al. (2003) Developmental Cell 5:367.
  4. Topol, L. et al. (2003) J. Cell Biol 162:899.
  5. Torres, M. et al. (1996) J. Cell Biol. 133:1123.
  6. Sen, M. et al. (2001) Arthritis & Rheumatism 44:772.

Long Name

Wingless-type MMTV Integration Site Family, Member 5a

Alternate Names

Wnt5a

Entrez Gene IDs

7474 (Human); 22418 (Mouse)

Gene Symbol

WNT5A

UniProt

Additional Wnt-5a Products

Product Documents for Human/Mouse Wnt-5a Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human/Mouse Wnt-5a Antibody

For research use only

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