Anti-Wnt3 antibody (ab32249)
Key features and details
- Rabbit polyclonal to Wnt3
- Suitable for: WB, IP, IHC-P
- Reacts with: Human, Recombinant fragment
- 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 -
Tested applications
Suitable for: WB, IP, IHC-Pmore details -
Species reactivity
Reacts with: Human, Recombinant fragment
Predicted to work with: Mouse, Rabbit, Chicken
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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
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
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
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 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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Western blot - Anti-Wnt3 antibody (ab32249)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. -
Immunoprecipitation - Anti-Wnt3 antibody (ab32249)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. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wnt3 antibody (ab32249)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
References (16)
ab32249 has been referenced in 16 publications.
- Wang J et al. (Pro)renin receptor promotes colorectal cancer through the Wnt/beta-catenin signalling pathway despite constitutive pathway component mutations. Br J Cancer 120:229-237 (2019). PubMed: 30555158
- Pouyafar A et al. Treatment of cancer stem cells from human colon adenocarcinoma cell line HT-29 with resveratrol and sulindac induced mesenchymal-endothelial transition rate. Cell Tissue Res N/A:N/A (2019). PubMed: 30758710
- Liu J et al. CircRNA_100367 regulated the radiation sensitivity of esophageal squamous cell carcinomas through miR-217/Wnt3 pathway. Aging (Albany NY) 11:12412-12427 (2019). PubMed: 31851619
- Song T et al. MicroRNA-2053 overexpression inhibits the development and progression of hepatocellular carcinoma. Oncol Lett 18:2043-2049 (2019). PubMed: 31423276
- Lan X et al. Nicotine enhances mesangial cell proliferation and fibronectin production in high glucose milieu via activation of Wnt/ß-catenin pathway. Biosci Rep 38:N/A (2018). PubMed: 29572389
Images
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Western blot - Anti-Wnt3 antibody (ab32249)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. -
Immunoprecipitation - Anti-Wnt3 antibody (ab32249)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. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Wnt3 antibody (ab32249)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.
