Validated using a knockout cell lineRecombinantRabMAb

Recombinant Alexa Fluor® 647 Anti-beta Catenin antibody [E247] (ab194119)

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Flow Cytometry - Alexa Fluor® 647 Anti-beta Catenin antibody [E247] (ab194119)
  • Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-beta Catenin antibody [E247] (ab194119)
  • Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-beta Catenin antibody [E247] (ab194119)
  • Flow Cytometry - Alexa Fluor® 647 Anti-beta Catenin antibody [E247] (ab194119)
  • Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-beta Catenin antibody [E247] (ab194119)
  • Alexa Fluor® 647 Anti-beta Catenin antibody [E247] (ab194119)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Alexa Fluor® 647 Rabbit monoclonal [E247] to beta Catenin
  • Suitable for: Flow Cyt, ICC/IF
  • Knockout validated
  • Reacts with: Human
  • Conjugation: Alexa Fluor® 647. Ex: 652nm, Em: 668nm

Overview

  • Product name

    Alexa Fluor® 647 Anti-beta Catenin antibody [E247]
    See all beta Catenin primary antibodies

  • Description

    Alexa Fluor® 647 Rabbit monoclonal [E247] to beta Catenin

  • Host species

    Rabbit

  • Conjugation

    Alexa Fluor® 647. Ex: 652nm, Em: 668nm

  • Tested applications

    Suitable for: Flow Cyt, ICC/IFmore details

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat

  • Immunogen

    Synthetic peptide within Human beta Catenin aa 1-100 (N terminal). The exact sequence is proprietary.

  • Positive control

    • ICC/IF: SW480 cells, Caco-2 cells, CTNNB1-HAP1 cells. Flow Cyt: SW480 cells.

  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    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, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab194119 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
Flow Cyt 1/500.

ab199093 - Rabbit monoclonal IgG (Alexa Fluor® 647), is suitable for use as an isotype control with this antibody.

 
ICC/IF 1/50 - 1/500.

Target

  • Function

    Key dowstream component of the canonical Wnt signaling pathway. 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. 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.
    Involved in the regulation of cell adhesion. The majority of beta-catenin is localized to the cell membrane and is part of E-cadherin/catenin adhesion complexes which are proposed to couple cadherins to the actin cytoskeleton.

  • Tissue specificity

    Expressed in several hair follicle cell types: basal and peripheral matrix cells, and cells of the outer and inner root sheaths. Expressed in colon.

  • Involvement in disease

    Defects in CTNNB1 are associated with colorectal cancer (CRC) [MIM:114500].
    Note=Activating mutations in CTNNB1 have oncogenic activity resulting in tumor development. Somatic mutations are found in various tumor types, including colon cancers, ovarian and prostate carcinomas, hepatoblastoma (HB), hepatocellular carcinoma (HCC). HBs are malignant embryonal tumors mainly affecting young children in the first three years of life.
    Defects in CTNNB1 are a cause of pilomatrixoma (PTR) [MIM:132600]; a common benign skin tumor.
    Defects in CTNNB1 are a cause of medulloblastoma (MDB) [MIM:155255]. MDB is a malignant, invasive embryonal tumor of the cerebellum with a preferential manifestation in children.
    Defects in CTNNB1 are a cause of susceptibility to ovarian cancer (OC) [MIM:167000]. Ovarian cancer common malignancy originating from ovarian tissue. Although many histologic types of ovarian neoplasms have been described, epithelial ovarian carcinoma is the most common form. Ovarian cancers are often asymptomatic and the recognized signs and symptoms, even of late-stage disease, are vague. Consequently, most patients are diagnosed with advanced disease.
    Note=A chromosomal aberration involving CTNNB1 is found in salivary gland pleiomorphic adenomas, the most common benign epithelial tumors of the salivary gland. Translocation t(3;8)(p21;q12) with PLAG1.

  • Sequence similarities

    Belongs to the beta-catenin family.
    Contains 12 ARM repeats.

  • Post-translational
    modifications

    Phosphorylation by GSK3B requires prior phosphorylation of Ser-45 by another kinase. Phosphorylation proceeds then from Thr-41 to Ser-37 and Ser-33.
    EGF stimulates tyrosine phosphorylation. Phosphorylation on Tyr-654 decreases CDH1 binding and enhances TBP binding.
    Ubiquitinated by the SCF(BTRC) E3 ligase complex when phosphorylated by GSK3B, leading to its degradation. Ubiquitinated by a E3 ubiquitin ligase complex containing UBE2D1, SIAH1, CACYBP/SIP, SKP1, APC and TBL1X, leading to its subsequent proteasomal degradation.

  • Cellular localization

    Cytoplasm. Nucleus. Cytoplasm > cytoskeleton. Cell junction > adherens junction. Cell junction. Cell membrane. Cytoplasmic when it is unstabilized (high level of phosphorylation) or bound to CDH1. Translocates to the nucleus when it is stabilized (low level of phosphorylation). Interaction with GLIS2 and MUC1 promotes nuclear translocation. Interaction with EMD inhibits nuclear localization.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • b-catenin antibody
    • Beta catenin antibody
    • Beta-catenin antibody
    • Cadherin associated protein antibody
    • Catenin (cadherin associated protein), beta 1, 88kDa antibody
    • Catenin beta 1 antibody
    • Catenin beta-1 antibody
    • CATNB antibody
    • CHBCAT antibody
    • CTNB1_HUMAN antibody
    • CTNNB antibody
    • CTNNB1 antibody
    • DKFZp686D02253 antibody
    • FLJ25606 antibody
    • FLJ37923 antibody
    • OTTHUMP00000162082 antibody
    • OTTHUMP00000165222 antibody
    • OTTHUMP00000165223 antibody
    • OTTHUMP00000209288 antibody
    • OTTHUMP00000209289 antibody
    see all

Images

  • Flow Cytometry - Alexa Fluor® 647 Anti-beta Catenin antibody [E247] (ab194119)
    Flow Cytometry - Alexa Fluor® 647 Anti-beta Catenin antibody [E247] (ab194119)

    Overlay histogram showing HAP1 wildtype (green line) and HAP1-CNNB1 knockout cells (red line) stained with ab194119. The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab194119, 0.01µg/ml) for 30 min at 22°C.

    A mouse IgG1 isotype control antibody  (ab199093) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-CNNB1 knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

    Acquisition of >5,000 events were collected using a 40 mW Red laser (640nm) and 670/14 bandpass filter.

    This antibody can also be used in HAP1 cells fixed with 4% formaldehyde (10 min) permeabilized with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

  • Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-beta Catenin antibody [E247] (ab194119)
    Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-beta Catenin antibody [E247] (ab194119)

    ab194119 staining β-catenin in wild-type HAP1 cells (top panel) and β-catenin knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab194119 at 1/500 dilution (shown in red) and ab195887 at 1/250 dilution (shown in green) overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-beta Catenin antibody [E247] (ab194119)
    Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-beta Catenin antibody [E247] (ab194119)

    ab194119 staining beta Catenin in Caco-2 cells. The cells were fixed with 4% formaldehyde (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab194119 at 1/100 dilution (shown in red) and ab195887, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 488), at 2µg/ml (shown in green). Nuclear DNA was labelled with DAPI (shown in blue).
    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Flow Cytometry - Alexa Fluor® 647 Anti-beta Catenin antibody [E247] (ab194119)
    Flow Cytometry - Alexa Fluor® 647 Anti-beta Catenin antibody [E247] (ab194119)
    Overlay histogram showing SW480 cells stained with ab194119 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab194119, 1/500 dilution) for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) Alexa Fluor® 647 used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a solid-state 25mW red diode laser (635 nm) and 675/30 bandpass filter.

    This antibody gave a positive signal in SW480 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
  • Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-beta Catenin antibody [E247] (ab194119)
    Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-beta Catenin antibody [E247] (ab194119)

    ab194119 staining beta Catenin in SW480 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilised in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab194119 at a working dilution of 1 in 50 (shown in red) and ab195887, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor® 488, shown in green) at 2µg/ml overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.

    This product gave a positive signal in 100% methanol (5 min) fixed SW480 cells under the same testing conditions.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Alexa Fluor® 647 Anti-beta Catenin antibody [E247] (ab194119)
    Alexa Fluor® 647 Anti-beta Catenin antibody [E247] (ab194119)

References (2)

Publishing research using ab194119? Please let us know so that we can cite the reference in this datasheet.

ab194119 has been referenced in 2 publications.

  • Polio SR  et al. Extracellular matrix stiffness regulates human airway smooth muscle contraction by altering the cell-cell coupling. Sci Rep 9:9564 (2019). PubMed: 31267003
  • Pägelow D  et al. The olfactory epithelium as a port of entry in neonatal neurolisteriosis. Nat Commun 9:4269 (2018). PubMed: 30323282

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