Human/Mouse/Rat beta-Catenin Antibody
R&D Systems | Catalog # AF1329
Key Product Details
Species Reactivity
Validated:
Human, Mouse, Rat
Cited:
Human, Mouse, Rat, Avian - Chicken, Hamster, Transgenic Mouse
Applications
Validated:
Immunohistochemistry, Western Blot, Intracellular Staining by Flow Cytometry, Immunocytochemistry, Simple Western, Chromatin Immunoprecipitation (ChIP), CyTOF-ready
Cited:
Immunohistochemistry, Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Immunocytochemistry, Immunoprecipitation, Chromatin Immunoprecipitation (ChIP), Chromatin Immunoprecipitation Sequencing, Bioassay, Functional Assay
Label
Unconjugated
Antibody Source
Polyclonal Goat IgG
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Product Specifications
Immunogen
E. coli-derived recombinant human beta -Catenin
Ala2-Leu781
Accession # P35222
Ala2-Leu781
Accession # P35222
Specificity
Detects human, mouse and rat beta -Catenin in Western blots.
Clonality
Polyclonal
Host
Goat
Isotype
IgG
Scientific Data Images for Human/Mouse/Rat beta-Catenin Antibody
Detection of Human/Mouse/Rat beta ‑Catenin by Western Blot.
Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line, C6 rat glioma cell line, and NIH-3T3 mouse embryonic fibroblast cell line. PVDF membrane was probed with 1 µg/mL Goat Anti-Human/Mouse/Rat beta -Catenin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1329) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). For additional reference, recombinant human beta -catenin (1 ng) was included. A specific band for beta-Catenin was detected at approximately 95 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.Detection of beta ‑Catenin-regul-ated Genes by Chromatin Immunoprecipitation.
HeLa human cervical epithelial carcinoma cell line were fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. beta -Catenin/ DNA complexes were immuno-precipitated using 5 µg Goat Anti-Human/Mouse/Rat beta -Catenin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1329) or control antibody (Catalog # AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (Catalog # BAF109). Immuno-complexes were captured using 50 µL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. TheSU(Z)12promoter was detected by standard PCR.beta ‑Catenin in SW480 Human Cell Line.
beta -Catenin was detected in immersion fixed SW480 human colorectal adenocarcinoma cell line using Goat Anti-Human/Mouse/Rat beta -Catenin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1329) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm and nuclei. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.beta -Catenin in Human Kidney Cancer Tissue.
beta -Catenin was detected in immersion fixed paraffin-embedded sections of human kidney cancer tissue using Goat Anti-Human/Mouse/Rat beta -Catenin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1329) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Lower panel shows a lack of labeling if primary antibodies are omitted and tissue is stained only with secondary antibody followed by incubation with detection reagents. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.beta ‑Catenin in Human Kidney Cancer Tissue.
beta -Catenin was detected in immersion fixed paraffin-embedded sections of human kidney cancer tissue using 15 µg/mL Goat Anti-Human/Mouse/Rat beta -Catenin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1329) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific labeling was localized to epithelial cells in collecting tubules in the medulla. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.Detection of beta ‑Catenin in HeLa Human Cell Line by Flow Cytometry.
HeLa human cervical epithelial carcinoma cell line was stained with Goat Anti-Human/Mouse/Rat beta -Catenin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1329, filled histogram) or control antibody (Catalog # AB-108-C, open histogram), followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0108). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.Detection of Human and Mouse beta ‑Catenin by Simple WesternTM.
Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line and NIH-3T3 mouse embryonic fibroblast cell line, loaded at 0.2 mg/mL. A specific band was detected for beta -Catenin at approximately 94-97 kDa (as indicated) using 50 µg/mL of Goat Anti-Human/Mouse/Rat beta -Catenin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1329) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system. Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.Detection of Human beta-Catenin by Western Blot
The effects of montelukast on ROS generation and of the inhibition of ROS generation on arsenic-induced EMT of NHBE cells. (A) Cells seeded in 96-well plates were pretreated with montelukast at 0.1 and 1 µM for 2 h, and then 1 µM NaAsO2 was applied for combination treatment. After 3 h of treatment, the cells were stained with 10 µM DCFH-DA for 30 min. After washing out the residue dye, the fluorescent signal was measured immediately. The quantitative results (left panel) and representative images (right panel) are shown. a: p < 0.05 compared with the vehicle control group. b: p < 0.05 compared with the NaAsO2 group. (B) Cells were pretreated with NAC for 2 h, and then 1 µM NaAsO2 for another 24 h. A wound was made with a 200-µl tip, and the wound coverage was analyzed 6 h after the wound was made. a: p < 0.05 compared with the vehicle control group. b: p < 0.05 compared with the NaAsO2 group. (C) Cells were pretreated with NAC for 2 h, and then 1 µM NaAsO2 was added for another 24 h. The cells were harvested for protein expression analysis by western blotting. a: p < 0.05 compared with the vehicle control group. b: p < 0.05 compared with the NaAsO2 group. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35517780), licensed under a CC-BY license. Not internally tested by R&D Systems.Detection of Human beta-Catenin by Western Blot
The effect of montelukast on arsenic-induced EMT marker expression in NHBE cells. Cells were pretreated with montelukast for 2 h, followed by combined treatment with 1 µM NaAsO2. After 24 h of treatment, the cells were harvested for protein expression analysis by western blotting. (A) Representative western blot images. (B–G) The quantitative results of protein expression changes are shown in (C–G). a: p < 0.05 compared with the vehicle control group. b: p < 0.05 compared with the NaAsO2 group. (H) The cell morphology changes. NHBE cells were pretreated with 0.1 or 1 µM montelukast respectively for 2 h and followed by sodium arsenite for another 48 h. The cell morphology was analyzed by radius-ratio methods. a: p < 0.05 compared with the vehicle control group. b: p < 0.05 compared with the NaAsO2 group. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35517780), licensed under a CC-BY license. Not internally tested by R&D Systems.Detection of Human beta-Catenin by Western Blot
Effect of the combination of montelukast and fluticasone on NaAsO2-induced cell migration of NHBE cells. Cells were pretreated with medications for 2 h and then arsenic was added. (A,B) After 24 h of incubation, the cells were trypsinized, plated onto the inner side of the Transwell chamber and incubated for 36 h. The migrated cells were analyzed by the ratio of the bottom/total of the cell occupied area. Representative images (left panel) and the quantitative results are shown (right panel). (C–I) After 24 of incubation, the protein lysates were extracted and applied to western blot analysis. The representative image was shown in (C), and the quantitative results were shown, respectively (D–I). a: p < 0.05 compared with the vehicle control group. b: p < 0.05 compared with the NaAsO2 group. c: p < 0.05 compared with the montelukast plus NaAsO2 group. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35517780), licensed under a CC-BY license. Not internally tested by R&D Systems.Detection of Human beta-Catenin by Western Blot
The effects of montelukast on NF-kappa B activation and of the inhibition of NF-kappa B on mediated arsenic-induced EMT of NHBE cells. (A) Arsenic induced the phosphorylation of AKT and p65. Cells were treated with 1 µM NaAsO2 for different time intervals, and the cells were harvested and subjected to western blot analysis using a phospho-antibody. (B) NAC treatment inhibited arsenic-induced phospho-p65. Cells were pretreated with NAC for 2 h, and then NaAsO2 was added for another 3 h. The expression of phospho-p65 was determined with a phospho-Ser536-p65 antibody. GAPDH served as the internal control. (C) Treatment with an NF-kappa B inhibitor reduced arsenic-induced cell migration. Cells were pretreated with 2 µM BAY117082 for 2 h and then treated with 1 µM NaAsO2 for another 24 h. A wound was made with a 200-µl tip, and the wound coverage was analyzed by comparing the cell-occupied area at 0 h versus that 6 h after the wound was made. The representative image (left panel) and the quantitative results (right panel) are shown. a: p < 0.05 compared with the vehicle control group. b: p < 0.05 compared with the NaAsO2 group. (D) Treatment with an NF-kappa B inhibitor reduced arsenic-induced mesenchymal marker expression. Cells were pretreated with 2 µM BAY117082 for 2 h, and then 1 µM NaAsO2 was added for another 24 h. The cells were harvested for protein expression analysis by western blotting. (E) Montelukast inhibited arsenic-induced phospho-p65 expression. Cells were pretreated with montelukast for 2 h, and then arsenic was added for another 3 h. The cells were harvested for protein expression analysis by western blotting. GAPDH served as the internal control. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35517780), licensed under a CC-BY license. Not internally tested by R&D Systems.Detection of beta-Catenin by Western Blot
Immunoblotting (a) with densitometry analysis, and (b) of the tdTomato-positive cells isolated from VE-cadherincreERT2RosatdTomatoIns2Akita/+ and VE-cadherincreERT2RosatdTomato mice treated with or without SB216763 (n = 3). The numbers represent each sample. Densitometry was analyzed for statistical significance by ANOVA with post hoc Tukey’s analysis. *** p < 0.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36983045), licensed under a CC-BY license. Not internally tested by R&D Systems.Detection of beta-Catenin by Western Blot
Immunoblotting (a) with densitometry analysis, and (b) of the tdTomato-positive cells isolated from VE-cadherincreERT2RosatdTomatoIns2Akita/+ and VE-cadherincreERT2RosatdTomato mice treated with or without SB216763 (n = 3). The numbers represent each sample. Densitometry was analyzed for statistical significance by ANOVA with post hoc Tukey’s analysis. *** p < 0.001. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/36983045), licensed under a CC-BY license. Not internally tested by R&D Systems.Applications for Human/Mouse/Rat beta-Catenin Antibody
Application
Recommended Usage
Chromatin Immunoprecipitation (ChIP)
5 µg/5 x 106 cells
Sample: HeLa human cervical epithelial carcinoma cell line chromatin, SU(Z)12 promoter detected by standard PCR.
Sample: HeLa human cervical epithelial carcinoma cell line chromatin, SU(Z)12 promoter detected by standard PCR.
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
Immunocytochemistry
5-15 µg/mL
Sample: Immersion fixed SW480 human colorectal adenocarcinoma cell line
Sample: Immersion fixed SW480 human colorectal adenocarcinoma cell line
Immunohistochemistry
5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human kidney cancer tissue
Sample: Immersion fixed paraffin-embedded sections of human kidney cancer tissue
Intracellular Staining by Flow Cytometry
2.5 µg/106 cells
Sample: HeLa human cervical epithelial carcinoma cell line fixed with paraformaldehyde and permeabilized with saponin
Sample: HeLa human cervical epithelial carcinoma cell line fixed with paraformaldehyde and permeabilized with saponin
Simple Western
50 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line and NIH‑3T3 mouse embryonic fibroblast cell line
Sample: HeLa human cervical epithelial carcinoma cell line and NIH‑3T3 mouse embryonic fibroblast cell line
Western Blot
1 µg/mL
Sample: HeLa human cervical epithelial carcinoma cell line, C6 rat glioma cell line, and NIH-3T3 mouse embryonic fibroblast cell line
Sample: HeLa human cervical epithelial carcinoma cell line, C6 rat glioma cell line, and NIH-3T3 mouse embryonic fibroblast cell line
Reviewed Applications
Read 6 reviews rated 4.3 using AF1329 in the following applications:
Flow Cytometry Panel Builder
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Advanced Features
- Spectra Viewer - Custom analysis of spectra from multiple fluorochromes
- Spillover Popups - Visualize the spectra of individual fluorochromes
- Antigen Density Selector - Match fluorochrome brightness with antigen density
Formulation, Preparation, and Storage
Purification
Antigen Affinity-purified
Reconstitution
Reconstitute at 0.2 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
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.
Calculators
Background: beta-Catenin
Alternate Names
bCatenin, CTNNB1
Gene Symbol
CTNNB1
UniProt
Additional beta-Catenin Products
Product Documents for Human/Mouse/Rat beta-Catenin Antibody
Certificate of Analysis
To download a Certificate of Analysis, please enter a lot or batch number in the search box below.
Note: Certificate of Analysis not available for kit components.
Product Specific Notices for Human/Mouse/Rat beta-Catenin Antibody
For research use only
Citations for Human/Mouse/Rat beta-Catenin Antibody
Customer Reviews for Human/Mouse/Rat beta-Catenin Antibody (6)
4.3 out of 5
6 Customer Ratings
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Application: ImmunohistochemistrySample Tested: epitheliumSpecies: MouseVerified Customer | Posted 06/27/2021
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Application: ImmunohistochemistrySample Tested: mouse oral epitheliumSpecies: MouseVerified Customer | Posted 11/23/2020
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Application: Immunocytochemistry/ImmunofluorescenceSample Tested: c2bbeSpecies: HumanVerified Customer | Posted 11/30/2018
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Application: Western BlotSample Tested: IPS2 induced pluripotent stem cellsSpecies: MouseVerified Customer | Posted 01/25/2018
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Application: ImmunofluorescenceSample Tested: kidneySpecies: RatVerified Customer | Posted 04/03/2015
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Application: ImmunoprecipitationSample Tested: See PMID 23474492Species: HumanVerified Customer | Posted 01/05/2015
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Protocols
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
- 7-Amino Actinomycin D (7-AAD) Cell Viability Flow Cytometry Protocol
- Antigen Retrieval Protocol (PIER)
- Antigen Retrieval for Frozen Sections Protocol
- Appropriate Fixation of IHC/ICC Samples
- Cellular Response to Hypoxia Protocols
- ChIP Protocol Video
- Chromatin Immunoprecipitation (ChIP) Protocol
- Chromatin Immunoprecipitation Protocol
- Chromogenic IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Chromogenic Immunohistochemistry Staining of Frozen Tissue
- ClariTSA™ Fluorophore Kits
- Detection & Visualization of Antibody Binding
- Extracellular Membrane Flow Cytometry Protocol
- Flow Cytometry Protocol for Cell Surface Markers
- Flow Cytometry Protocol for Staining Membrane Associated Proteins
- Flow Cytometry Staining Protocols
- Flow Cytometry Troubleshooting Guide
- Fluorescent IHC Staining of Frozen Tissue Protocol
- Graphic Protocol for Heat-induced Epitope Retrieval
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Graphic Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Graphic Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- ICC Cell Smear Protocol for Suspension Cells
- ICC Immunocytochemistry Protocol Videos
- ICC for Adherent Cells
- IHC Sample Preparation (Frozen sections vs Paraffin)
- Immunocytochemistry (ICC) Protocol
- Immunocytochemistry Troubleshooting
- Immunofluorescence of Organoids Embedded in Cultrex Basement Membrane Extract
- Immunofluorescent IHC Staining of Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Protocol
- Immunohistochemistry (IHC) and Immunocytochemistry (ICC) Protocols
- Immunohistochemistry Frozen Troubleshooting
- Immunohistochemistry Paraffin Troubleshooting
- Intracellular Flow Cytometry Protocol Using Alcohol (Methanol)
- Intracellular Flow Cytometry Protocol Using Detergents
- Intracellular Nuclear Staining Flow Cytometry Protocol Using Detergents
- Intracellular Staining Flow Cytometry Protocol Using Alcohol Permeabilization
- Intracellular Staining Flow Cytometry Protocol Using Detergents to Permeabilize Cells
- Preparing Samples for IHC/ICC Experiments
- Preventing Non-Specific Staining (Non-Specific Binding)
- Primary Antibody Selection & Optimization
- Propidium Iodide Cell Viability Flow Cytometry Protocol
- Protocol for Heat-Induced Epitope Retrieval (HIER)
- Protocol for Liperfluo
- Protocol for Making a 4% Formaldehyde Solution in PBS
- Protocol for VisUCyte™ HRP Polymer Detection Reagent
- Protocol for the Characterization of Human Th22 Cells
- Protocol for the Characterization of Human Th9 Cells
- Protocol for the Fluorescent ICC Staining of Cell Smears - Graphic
- Protocol for the Fluorescent ICC Staining of Cultured Cells on Coverslips - Graphic
- Protocol for the Preparation & Fixation of Cells on Coverslips
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Frozen Tissue Sections - Graphic
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation and Chromogenic IHC Staining of Paraffin-embedded Tissue Sections - Graphic
- Protocol for the Preparation and Fluorescent ICC Staining of Cells on Coverslips
- Protocol for the Preparation and Fluorescent ICC Staining of Non-adherent Cells
- Protocol for the Preparation and Fluorescent ICC Staining of Stem Cells on Coverslips
- Protocol for the Preparation and Fluorescent IHC Staining of Frozen Tissue Sections
- Protocol for the Preparation and Fluorescent IHC Staining of Paraffin-embedded Tissue Sections
- Protocol for the Preparation of Gelatin-coated Slides for Histological Tissue Sections
- Protocol for the Preparation of a Cell Smear for Non-adherent Cell ICC - Graphic
- Protocol: Annexin V and PI Staining by Flow Cytometry
- Protocol: Annexin V and PI Staining for Apoptosis by Flow Cytometry
- R&D Systems Quality Control Western Blot Protocol
- TUNEL and Active Caspase-3 Detection by IHC/ICC Protocol
- The Importance of IHC/ICC Controls
- Troubleshooting Guide: Fluorokine Flow Cytometry Kits
- Troubleshooting Guide: Immunohistochemistry
- Troubleshooting Guide: Western Blot Figures
- Western Blot Conditions
- Western Blot Protocol
- Western Blot Protocol for Cell Lysates
- Western Blot Troubleshooting
- Western Blot Troubleshooting Guide
- View all Protocols, Troubleshooting, Illustrated assays and Webinars
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Associated Pathways
Blood-Brain Barrier Pathway: Anatomy
HIF Enhancer Pathways
Notch Signaling Pathways
Wnt Signaling Pathways: beta-Catenin-dependent Wnt Signaling