Product nameAnti-Wnt3 antibody
See all Wnt3 primary antibodies
DescriptionRabbit polyclonal to Wnt3
Tested applicationsSuitable for: WB, IP, IHC-Pmore details
Species reactivityReacts with: Mouse, Human
Predicted to work with: Rabbit, Chicken
- A549 (Human Lung Carcinoma) Whole Cell Lysate Human lung adenocarcinoma FFPE tissue sections
Storage instructionsShipped 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 bufferPreservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration information loading...
PurityImmunogen affinity purified
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.
|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.|
FunctionLigand 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 diseaseDefects 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 similaritiesBelongs to the Wnt family.
Cellular localizationSecreted > extracellular space > extracellular matrix.
- Information by UniProt
- Int 4 antibody
- INT4 antibody
- MGC131950 antibody
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.
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
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.
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.
This product has been referenced in:
- Sun X et al. Colorectal cancer cells suppress CD4+ T cells immunity through canonical Wnt signaling. Oncotarget 8:15168-15181 (2017). Read more (PubMed: 28147310) »
- Richards MH et al. Dynamic interaction between astrocytes and infiltrating PBMCs in context of neuroAIDS. Glia 63:441-51 (2015). Read more (PubMed: 25331637) »