Knockout Tested Rabbit Recombinant Monoclonal beta Catenin antibody. Carrier free. Suitable for WB, IHC-P, ICC/IF, Flow Cyt (Intra), IP, Dot and reacts with Human, Mouse, Rat samples.
pH: 7.2 - 7.4
Constituents: 100% PBS
WB | IHC-P | ICC/IF | Flow Cyt (Intra) | IP | Dot | |
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Human | Tested | Tested | Tested | Tested | Tested | Tested |
Mouse | Tested | Tested | Tested | Tested | Tested | Expected |
Rat | Tested | Tested | Expected | Expected | Expected | Expected |
Species | Dilution info | Notes |
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Species Human, Mouse, Rat | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Mouse | Dilution info - | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species Rat | Dilution info - | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
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Species Human, Mouse | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
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Species Human, Mouse | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
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Species Human, Mouse | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Mouse, Rat | Dilution info Use at an assay dependent concentration. | Notes - |
Key downstream component of the canonical Wnt signaling pathway (PubMed:17524503, PubMed:18077326, PubMed:18086858, PubMed:18957423, PubMed:21262353, PubMed:22155184, PubMed:22647378, PubMed:22699938). In the absence of Wnt, forms a complex with AXIN1, AXIN2, APC, CSNK1A1 and GSK3B that promotes phosphorylation on N-terminal Ser and Thr residues and ubiquitination of CTNNB1 via BTRC and its subsequent degradation by the proteasome (PubMed:17524503, PubMed:18077326, PubMed:18086858, PubMed:18957423, PubMed:21262353, PubMed:22155184, PubMed:22647378, PubMed:22699938). In the presence of Wnt ligand, CTNNB1 is not ubiquitinated and accumulates in the nucleus, where it acts as a coactivator for transcription factors of the TCF/LEF family, leading to activate Wnt responsive genes (PubMed:17524503, PubMed:18077326, PubMed:18086858, PubMed:18957423, PubMed:21262353, PubMed:22155184, PubMed:22647378, PubMed:22699938). Involved in the regulation of cell adhesion, as component of an E-cadherin:catenin adhesion complex (By similarity). Acts as a negative regulator of centrosome cohesion (PubMed:18086858). Involved in the CDK2/PTPN6/CTNNB1/CEACAM1 pathway of insulin internalization (PubMed:21262353). Blocks anoikis of malignant kidney and intestinal epithelial cells and promotes their anchorage-independent growth by down-regulating DAPK2 (PubMed:18957423). Disrupts PML function and PML-NB formation by inhibiting RANBP2-mediated sumoylation of PML (PubMed:22155184). Promotes neurogenesis by maintaining sympathetic neuroblasts within the cell cycle (By similarity). Involved in chondrocyte differentiation via interaction with SOX9: SOX9-binding competes with the binding sites of TCF/LEF within CTNNB1, thereby inhibiting the Wnt signaling (By similarity). Acts as a positive regulator of odontoblast differentiation during mesenchymal tooth germ formation, via promoting the transcription of differentiation factors such as LEF1, BMP2 and BMP4 (By similarity). Activity is repressed in a MSX1-mediated manner at the bell stage of mesenchymal tooth germ formation which prevents premature differentiation of odontoblasts (By similarity).
CTNNB, OK/SW-cl.35, PRO2286, CTNNB1, Catenin beta-1, Beta-catenin
Knockout Tested Rabbit Recombinant Monoclonal beta Catenin antibody. Carrier free. Suitable for WB, IHC-P, ICC/IF, Flow Cyt (Intra), IP, Dot and reacts with Human, Mouse, Rat samples.
pH: 7.2 - 7.4
Constituents: 100% PBS
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
Beta Catenin also known by names such as CTNNB1 or beta-chip is an important protein involved in cell signaling and adhesion. This protein has a molecular weight of around 88 kDa. Beta Catenin is expressed in many cell types and tissues indicating its widespread role in various biological processes. It functions mechanically by mediating the linkage between cadherins and the actin cytoskeleton facilitating cell-cell adhesion. Beta Catenin is also a central part of transcription regulation processes in the nucleus.
This protein plays roles in both cell adhesion and the regulation of gene expression. Beta Catenin is a critical component of the Wnt signaling pathway where it can form complexes with other proteins to influence gene transcription. In the absence of Wnt signaling beta Catenin levels are low due to its degradation. However when the pathway is active it accumulates in the cytoplasm and eventually translocates to the nucleus where it interacts with TCF/LEF transcription factors to regulate the expression of target genes.
Beta Catenin plays a central role in the Wnt signaling pathway and influences cell fate decisions and cellular proliferation. It acts in concert with proteins such as Dishevelled (DVL) and Axin to coordinate these important biological processes. In the absence of Wnt signaling proteins such as APC and GSK-3β are responsible for beta Catenin degradation keeping its cellular levels in check. Beta Catenin’s interaction with transcription factors in the nucleus makes it pivotal in the regulation of cell and tissue homeostasis.
Beta Catenin has associations with colorectal cancer and hepatocellular carcinoma. Its dysregulation can lead to unchecked cell proliferation and tumorigenesis. Often mutations in the beta Catenin gene (CTNNB1) or components of the Wnt pathway like APC are implicated in the development of these cancers. Its interplay with E-cadherin is important for maintaining tissue architecture and disruptions can lead to invasive cancer phenotypes. Understanding beta Catenin’s role provides insights into therapeutic strategies for these cancers.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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This data was developed using Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] ab305261, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling beta Catenin non-phospho(active) S45 with Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] ab305261 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing membranous and weak cytoplasmic staining in NIH/3T3 cell line. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8)
Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red).
The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.
This data was developed using Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] ab305261, the same antibody clone in a different buffer formulation.
Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized CTNNB1 (beta Catenin) knockout HAP1 cells labelling beta Catenin non-phospho (active) S45 with Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] ab305261 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing membranous staining in parental HAP1 cell line, and no staining in CTNNB1 (beta Catenin) knockout HAP1 cell line. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8)
Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).
Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.
This data was developed using Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] ab305261, the same antibody clone in a different buffer formulation.
beta Catenin non-phospho(active) S45 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 μg with Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] ab305261 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] ab305261 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate 10 μg
Lane 2: Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] ab305261 IP in NIH/3T3 whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] ab305261 in NIH/3T3 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 50 seconds
All lanes: Immunoprecipitation - Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] (Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] ab305261) at 1/30 dilution
All lanes: HeLa (human cervix adenocarcinoma epithelial cell)
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Observed band size: 95 kDa
Exposure time: 50s
This data was developed using Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] ab305261, the same antibody clone in a different buffer formulation.
beta Catenin non-phospho(active) S45 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 μg with Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] ab305261 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] ab305261 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(VeriBlot for IP Detection Reagent (HRP) ab131366) was used at 1/5000 dilution.
Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 μg
Lane 2: Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] ab305261 IP in HeLa whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] ab305261 in HeLa whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 50 seconds
All lanes: Immunoprecipitation - Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] (Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] ab305261) at 1/30 dilution
All lanes: HeLa (human cervix adenocarcinoma epithelial cell)
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Observed band size: 95 kDa
Exposure time: 50s
Lane 1: beta Catenin non-phospho peptide
Lane 2: beta Catenin phospho (S33) peptide
Lane 3: beta Catenin phospho (S33/S37) peptide
Lane 4: beta Catenin phospho (S33/S37/T41) peptide
Lane 5: beta Catenin phospho (S37) peptide
Lane 6: beta Catenin phospho (S37/T41) peptide
Lane 7: beta Catenin phospho (T41) peptide
Lane 8: beta Catenin phospho (S33/T41) peptide
Lane 9: beta Catenin phospho (S33/S37/T41/S45) peptide
Lane 10: beta Catenin phospho (S37/T41/S45) peptide
Lane 11: beta Catenin phospho (T41/S45) peptide
Lane 12: beta Catenin phospho (S45) peptide
Lane 13: beta Catenin phospho (S33/T41/S45) peptide
Lane 14: beta Catenin phospho (S33/S45) peptide
Lane 15: beta Catenin phospho (S37/S45) peptide
This data was developed using Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] ab305261, the same antibody clone in a different buffer formulation.
Dot blot analysis of beta Catenin non-phospho (active) S45 using Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] ab305261 at 1/1000 dilution (0.454 µg/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100,000 dilution.
Exposure time: 180 seconds
Blocking and diluting buffer and concentration: 5% NFDM/TBST
This data was developed using Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] ab305261, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration:Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS The samples were run on a Bis-Tris gel. Performed under reducing conditions. False colour image of Western blot: Anti-beta Catenin non-phospho(active) S45 antibody (Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] ab305261) staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody 6C5 (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] ab305261 was shown to bind specifically to beta Catenin non-phospho(active) S45. A band was observed at 95 kDa in wild-type HAP1 cell lysates with no signal observed at this size in CTNNB1 (beta Catenin) knockout cell line. To generate this image, wild-type and CTNNB1 (beta Catenin) knockout HAP1 cell lysates were analyzed. First, samples were run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in in Intercept® (TBS) Blocking Buffer diluted with an equal volume of 0.1% TBS before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] (Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] ab305261) at 1/1000 dilution
Lane 1: Wild-type HAP1(human chronic myelogenous leukemia near-haploid cell line) whole cell lysate at 20 µg
Lane 2: CTNNB1 (beta Catenin) knockout HAP1 whole cell lysate at 20 µg
Lanes 1 - 2: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) at 1/10000 dilution
Lanes 1 - 2: Western blot - Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/10000 dilution
Observed band size: 95 kDa
This data was developed using Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] ab305261, the same antibody clone in a different buffer formulation.
Blocking and diluting buffer and concentration: 5% NFDM/TBST
Lysates were freshly made and used for Western Blotting immediately to minimize protein degradation.
Exposure time: 180 seconds
All lanes: Western blot - Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] (Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] ab305261) at 1/1000 dilution
Lane 1: HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg
Lane 2: 293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg
Lane 3: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg
Lane 4: PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/50000 dilution
Observed band size: 95 kDa
Exposure time: 180s
This data was developed using Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] ab305261, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell, Right) / CTNNB1 (beta Catenin) knockout HAP1(Left) cells labelling beta Catenin non-phospho(active) S45 with Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] ab305261 at 1/500 dilution (0.1ug) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody.
This data was developed using Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] ab305261, the same antibody clone in a different buffer formulation.
Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling beta Catenin non-phospho(active) S45 with Anti-beta Catenin non-phospho(active) S45 antibody [EPR26155-110-1] ab305261 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 1/2000 dilution was used as the secondary antibody.
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