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  1. Link

    beta-catenin-antibody-igx4794r-3-ab223075.pdf

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Signal Transduction Cytoskeleton / ECM Cell Adhesion Cadherins
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Validated using a knockout cell lineRecombinant

Recombinant Anti-beta Catenin antibody [IGX4794R-3] (ab223075)

  • Datasheet
  • SDS
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Western blot - Anti-beta Catenin antibody [IGX4794R-3] (ab223075)
  • Western blot - Anti-beta Catenin antibody [IGX4794R-3] (ab223075)
  • Immunocytochemistry/ Immunofluorescence - Anti-beta Catenin antibody [IGX4794R-3] (ab223075)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [IGX4794R-3] (ab223075)
  • Western blot - Anti-beta Catenin antibody [IGX4794R-3] (ab223075)
  • Western blot - Anti-beta Catenin antibody [IGX4794R-3] (ab223075)
  • Immunocytochemistry/ Immunofluorescence - Anti-beta Catenin antibody [IGX4794R-3] (ab223075)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [IGX4794R-3] (ab223075)
  • Anti-beta Catenin antibody [IGX4794R-3] (ab223075)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [IGX4794R-3] to beta Catenin
  • Suitable for: WB, ICC/IF, IHC-Fr, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Human

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Overview

  • Product name

    Anti-beta Catenin antibody [IGX4794R-3]
    See all beta Catenin primary antibodies
  • Description

    Rabbit monoclonal [IGX4794R-3] to beta Catenin
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, IHC-Fr, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HCT116, A549, Caco2, Hues7, NIH3T3, mES, E14Tg2a and wildtype HAP1 whole cell and human colon tissue lysates. IHC-P: FFPE Human Colon (Normal) and Human Colon Adenocarcinoma tissue sections. ICC/IF: MCF-7 and wildtype HAP1 cells.
  • General notes

    This product was made using synthetic libraries and phage display technology.

    This antibody is a recombinant chimeric antibody. Rabbit chimeric monoclonal antibody (Human Fab/ Rabbit Fc).

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.02% Sodium azide
    Constituents: 1% BSA, PBS
  • Concentration information loading...
  • Clonality

    Monoclonal
  • Clone number

    IGX4794R-3
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Cytoskeleton / ECM
    • Cell Adhesion
    • Cadherins
    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Microfilaments
    • Actin etc
    • Catenins
    • Neuroscience
    • Neurology process
    • Neurogenesis
    • Stem Cells
    • Signaling Pathways
    • Wnt
    • Cytoplasmic
    • Cancer
    • Invasion/microenvironment
    • ECM
    • Cell adhesion
    • Cadherins
    • Cardiovascular
    • Atherosclerosis
    • Vascular Inflammation
    • Leukocyte recruitment
    • Other

Associated products

  • KO cell lines

    • Human CTNNB1 (beta Catenin II) knockout HCT116 cell line (ab273712)
  • KO cell lysates

    • Human CTNNB1 (beta Catenin) knockout HCT116 cell lysate (ab275247)
  • Recombinant Protein

    • Recombinant Human beta Catenin protein (Tagged) (ab63175)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab223075 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 92 kDa (predicted molecular weight: 85 kDa).
ICC/IF
Use a concentration of 0.05 µg/ml.
IHC-Fr (1)
Use at an assay dependent concentration.
IHC-P
Use a concentration of 0.25 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Notes
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 92 kDa (predicted molecular weight: 85 kDa).
ICC/IF
Use a concentration of 0.05 µg/ml.
IHC-Fr
Use at an assay dependent concentration.
IHC-P
Use a concentration of 0.25 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

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.
  • Target information above from: UniProt accession P35222 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 1499 Human
    • Entrez Gene: 12387 Mouse
    • Omim: 116806 Human
    • SwissProt: P35222 Human
    • SwissProt: Q02248 Mouse
    • Unigene: 476018 Human
    • Unigene: 291928 Mouse
    • 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

    • Western blot - Anti-beta Catenin antibody [IGX4794R-3] (ab223075)
      Western blot - Anti-beta Catenin antibody [IGX4794R-3] (ab223075)
      All lanes : Anti-beta Catenin antibody [IGX4794R-3] (ab223075) at 1 µg/ml

      Lane 1 : Wild-type HCT 116 cell lysate
      Lane 2 : CTNNB1 CRISPR-Cas9 edited HCT 116 cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 85 kDa
      Observed band size: 95 kDa why is the actual band size different from the predicted?



      False colour image of Western blot: Anti-beta Catenin antibody [IGX4794R-3] staining at 1 ug/ml, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab223075 was shown to bind specifically to beta Catenin. A band was observed at 95 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in CTNNB1 CRISPR-Cas9 edited cell line ab273712 (CRISPR-Cas9 edited cell lysate ab275247). The band observed in the CRISPR-Cas9 edited lysate lane below 95 kDa is likely to represent a truncated form of beta Catenin. This has not been investigated further and the functional properties of the gene product have not been determined. To generate this image, wild-type and CTNNB1 CRISPR-Cas9 edited HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

    • Western blot - Anti-beta Catenin antibody [IGX4794R-3] (ab223075)
      Western blot - Anti-beta Catenin antibody [IGX4794R-3] (ab223075)
      All lanes : Anti-beta Catenin antibody [IGX4794R-3] (ab223075) at 1 µg/ml

      Lane 1 : Wild-type HCT116 cell lysate
      Lane 2 : CTNNB1 knockout HCT116 cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 85 kDa
      Observed band size: 95 kDa why is the actual band size different from the predicted?



      Lanes 1 - 2: Merged signal (red and green). Green - ab223075 observed at 95 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

      ab223075 was shown to react with Anti-beta Catenin in HCT 116 wild-type cells in western blot with loss of signal observed in CTNNB1 knockout cell line ab273712 (CTNNB1 knockout cell lysate ab275247). HCT 116 wild-type and CTNNB1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluoroscent western blot (TBS-based) blocking solution 50% (v/v) in TBS-T (0.1% Tween®) before incubation with ab223075 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at 1 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-beta Catenin antibody [IGX4794R-3] (ab223075)
      Immunocytochemistry/ Immunofluorescence - Anti-beta Catenin antibody [IGX4794R-3] (ab223075)

      ab223075 staining β-Catenin in wild-type HAP1 cells (top panel) and CTNNB1 (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 ab223075 at 0.05μg/ml and ab195889 at 1/250 dilution (shown in red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor 488) (ab150081) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [IGX4794R-3] (ab223075)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [IGX4794R-3] (ab223075)

      IHC image of CTNNB1 staining in a section of formalin-fixed paraffin-embedded human colon (normal)*, performed on a Leica BondTM. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins, before blocking of endogenous biotin using ab64212. The section was then incubated with ab223075, 0.25ug/ml, for 15 mins at room temperature and detected using an HRP conjugated ABC system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.

      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.

      *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

    • Western blot - Anti-beta Catenin antibody [IGX4794R-3] (ab223075)
      Western blot - Anti-beta Catenin antibody [IGX4794R-3] (ab223075)
      All lanes : Anti-beta Catenin antibody [IGX4794R-3] (ab223075) at 1 µg/ml

      Lane 1 : A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate
      Lane 2 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate
      Lane 3 : HUES7 (Human embryonic stem cell line) Whole Cell Lysate
      Lane 4 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
      Lane 5 : mES (Mouse embryonic stem cell) Whole Cell Lysate
      Lane 6 : E14Tg2a (Mouse embryonic stem cell line) Whole Cell Lysate
      Lane 7 : Colon (Human) Tissue Lysate - adult normal tissue
      Lane 8 : Wild type HAP1 whole cell lysate
      Lane 9 : Beta Catenin knockout HAP1 whole cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Peroxidase AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/50000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 85 kDa
      Observed band size: 92 kDa why is the actual band size different from the predicted?


      Exposure time: 3 minutes


      This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab223075 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

    • Western blot - Anti-beta Catenin antibody [IGX4794R-3] (ab223075)
      Western blot - Anti-beta Catenin antibody [IGX4794R-3] (ab223075)

      Lane 1: Wild type HAP1 whole cell lysate (10 µg)
      Lane 2: CTNNB1 (β-Catenin) knockout HAP1 whole cell lysate (10 µg)
      Lane 3: A431 whole cell lysate (10 µg)
      Lane 4: Caco-2 whole cell lysate (10 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab223075 observed at 95 kDa. Red - loading control, ab9484, observed at 37 kDa.

      ab223075 was shown to specifically react with CTNNB1 (β-Catenin) in wild-type HAP1 cells along with additional cross reactive bands. No band was observed when knockout samples were used. Wild-type and CTNNB1 (β-Catenin) knockout samples were subjected to SDS-PAGE. ab223075 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1 µg/ml and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-beta Catenin antibody [IGX4794R-3] (ab223075)
      Immunocytochemistry/ Immunofluorescence - Anti-beta Catenin antibody [IGX4794R-3] (ab223075)

      ab223075 staining CTNNB1 in MCF-7 cells. The cells were fixed with 100% methanol (5 min), 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 ab223075 at a 0.05µg/ml concentration, then detected with an Alexa Fluor® 488 goat anti-rabbit secondary antibody (ab150081) at a 1/1000 dilution (shown in green). Nuclear DNA was labelled with DAPI (shown in blue), and ab195889, Mouse monoclonal to alpha Tubulin (Alexa Fluor® 594), at a 1/250 dilution (shown in red).

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

      This product also gave a positive signal in 4% formaldehyde (10 min) fixed MCF-7 cells under the same testing conditions.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [IGX4794R-3] (ab223075)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta Catenin antibody [IGX4794R-3] (ab223075)

      IHC image of CTNNB1 staining in a section of formalin-fixed paraffin-embedded human colon adenocarcinoma*, performed on a Leica BONDTM. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins.  The section was then incubated with ab223075, 0.1ug/ml dilution, 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. The inset negative control image is taken from an identical assay without primary antibody.

      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.

      *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

    • Anti-beta Catenin antibody [IGX4794R-3] (ab223075)
      Anti-beta Catenin antibody [IGX4794R-3] (ab223075)

    Protocols

    • Western blot protocols
    • Immunohistochemistry protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (9)

    Publishing research using ab223075? Please let us know so that we can cite the reference in this datasheet.

    ab223075 has been referenced in 9 publications.

    • Wang Z  et al. High-throughput sequencing profile of laryngeal cancers: analysis of co-expression and competing endogenous RNA networks of circular RNAs, long non-coding RNAs, and messenger RNAs. Ann Transl Med 9:483 (2021). PubMed: 33850880
    • Wang Z  et al. Downregulation of circ-RANBP9 in laryngeal cancer and its clinical significance. Ann Transl Med 9:484 (2021). PubMed: 33850881
    • Liang G  et al. Overexpression of hsa_circ_0001445 reverses oxLDL-induced inhibition of HUVEC proliferation via SRSF1. Mol Med Rep 24:N/A (2021). PubMed: 33982782
    • Bai J  et al. Glycyrrhizic Acid Promotes Osteogenic Differentiation of Human Bone Marrow Stromal Cells by Activating the Wnt/ß-Catenin Signaling Pathway. Front Pharmacol 12:607635 (2021). PubMed: 33935702
    • Liu B  et al. Exosome-derived microRNA-433 inhibits tumorigenesis through incremental infiltration of CD4 and CD8 cells in non-small cell lung cancer. Oncol Lett 22:607 (2021). PubMed: 34188709
    • Hyun J  et al. Morus alba Root Extract Induces the Anagen Phase in the Human Hair Follicle Dermal Papilla Cells. Pharmaceutics 13:N/A (2021). PubMed: 34452116
    • Silva C  et al. Selective pro-apoptotic and antimigratory effects of polyphenol complex catechin:lysine 1:2 in breast, pancreatic and colorectal cancer cell lines. Eur J Pharmacol 859:172533 (2019). PubMed: 31301308
    • Hoyt AK  et al. PRP-1 significantly decreases the ALDHhigh cancer stem cell population and regulates the aberrant Wnt/ß-catenin pathway in human chondrosarcoma JJ012 cells. Oncol Rep 42:103-114 (2019). PubMed: 31180539
    • Deng B  et al. Exendin-4 promotes osteogenic differentiation of adipose-derived stem cells and facilitates bone repair. Mol Med Rep 20:4933-4942 (2019). PubMed: 31661134

    Customer reviews and Q&As

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    Immunohistochemistry (Frozen sections) abreview for Anti-beta Catenin antibody [IGX4794R-3]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Frozen sections)
    Sample
    Mouse Tissue sections (skin)
    Permeabilization
    Yes - 0.05% Tween
    Specification
    skin
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    Radhika Atit

    Verified customer

    Submitted Nov 29 2021

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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