Anti-Wnt3 antibody (ab32249)
Key features and details
- Rabbit polyclonal to Wnt3
- Suitable for: WB, IP, IHC-P
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-Wnt3 antibody
See all Wnt3 primary antibodies -
Description
Rabbit polyclonal to Wnt3 -
Host species
Rabbit -
Specificity
From Jan 2024, QC testing of replenishment batches of this polyclonal changed. All tested and expected application and reactive species combinations are still covered by our Abcam product promise. However, we no longer test all applications. For more information on a specific batch, please contact our Scientific Support who will be happy to help.
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Tested applications
Suitable for: WB, IP, IHC-Pmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rabbit, Chicken -
Immunogen
Synthetic peptide corresponding to Mouse Wnt3 aa 250-350 conjugated to keyhole limpet haemocyanin.
(Peptide available asab32248) -
Positive control
- A549 (Human Lung Carcinoma) Whole Cell Lysate Human lung adenocarcinoma FFPE tissue sections
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help. -
Concentration information loading...
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Purity
Immunogen affinity purified -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab32249 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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WB |
Use a concentration of 1 µg/ml. Detects a band of approximately 37 kDa (predicted molecular weight: 40 kDa).
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IP |
Use at an assay dependent concentration.
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IHC-P |
Use at an assay dependent concentration.
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Notes |
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WB
Use a concentration of 1 µg/ml. Detects a band of approximately 37 kDa (predicted molecular weight: 40 kDa). |
IP
Use at an assay dependent concentration. |
IHC-P
Use at an assay dependent concentration. |
Target
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Function
Ligand for members of the frizzled family of seven transmembrane receptors. Wnt-3 and Wnt-3a play distinct roles in cell-cell signaling during morphogenesis of the developing neural tube. -
Involvement in disease
Defects in WNT3 are the cause of autosomal recessive tetra-amelia (ARTTRA) [MIM:273395]. Tetra-amelia is a rare human genetic disorder characterized by complete absence of all four limbs and other anomalies such as craniofacial, nervous system, pulmonary, skeletal and urogenital defects. -
Sequence similarities
Belongs to the Wnt family. -
Cellular localization
Secreted > extracellular space > extracellular matrix. - Information by UniProt
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Database links
- Entrez Gene: 374142 Chicken
- Entrez Gene: 7473 Human
- Entrez Gene: 22415 Mouse
- Omim: 165330 Human
- SwissProt: P56703 Human
- SwissProt: P17553 Mouse
- Unigene: 445884 Human
- Unigene: 159091 Mouse
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Alternative names
- Int 4 antibody
- INT4 antibody
- MGC131950 antibody
see all
Images
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All lanes : Anti-Wnt3 antibody (ab32249) at 1 µg/ml
Lane 1 : MCF-7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µg
Lane 2 : A549 (Human lung carcinoma cell line) Whole Cell Lysate at 10 µg
Lane 3 : Wnt3a Recombinant Protein at 0.01 µg
Secondary
All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 40 kDa
Observed band size: 37 kDa why is the actual band size different from the predicted?
Additional bands at: 50 kDa (possible cross reactivity)
ab32249 is not expected to recognize Wnt3a due to the immunogen having low sequence homology. -
Wnt3 was immunoprecipitated using 0.5mg MCF 7 whole cell extract, 5µg of Rabbit polyclonal to Wnt3 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
The antibody was incubated under agitation with Protein G beads for 10min, MCF 7 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab32249.
Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).
Band: 40kDa: Wnt3. -
IHC image of ab32249 staining in human lung adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab32249, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (23)
ab32249 has been referenced in 23 publications.
- Wang J et al. (Pro)renin receptor promotes colorectal cancer progression through inhibiting the NEDD4L-mediated Wnt3 ubiquitination and modulating gut microbiota. Cell Commun Signal 21:2 (2023). PubMed: 36597142
- Li Y et al. LincRNA-EPS increases TGF-β expression to inhibit the Wnt/β-catenin pathway, VSMC osteoblastic differentiation and vascular calcification in diabetic mice. Exp Ther Med 23:425 (2022). PubMed: 35607373
- Xu W et al. Molecular subtypes based on Wnt-signaling gene expression predict prognosis and tumor microenvironment in hepatocellular carcinoma. Front Immunol 13:1010554 (2022). PubMed: 36275697
- Cho YH et al. 5-FU promotes stemness of colorectal cancer via p53-mediated WNT/ß-catenin pathway activation. Nat Commun 11:5321 (2020). PubMed: 33087710
- Tanioka M et al. Lipid Emulsion Improves Functional Recovery in an Animal Model of Stroke. Int J Mol Sci 21:N/A (2020). PubMed: 33036206