Validated using a knockout cell line
Recombinant
RabMAb

Anti-beta Catenin antibody [E247] (ab32572)

Overview

  • Product name
    Anti-beta Catenin antibody [E247]
    See all beta Catenin primary antibodies
  • Description
    Rabbit monoclonal [E247] to beta Catenin
  • Tested applications
    Suitable for: IHC-Fr, IHC-P, WB, ICC/IF, IPmore details
    Unsuitable for: Flow Cyt
  • Species reactivity
    Reacts with: Mouse, Rat, Sheep, Hamster, Cow, Human, Macaque monkey, African green monkey
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human beta Catenin aa 1-100 (N terminal).

  • Positive control
    • WB: A431 cell lysate. ICC/IF: A431 and wildtype HAP1 cells. IHC-P: Human colon adenocarcinoma tissue.
  • General notes

    A trial size is available to purchase for this antibody.

    Alternative versions available:

    Anti-beta Catenin antibody (BSA & Azide free) [E247] (ab196204)

    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 rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab32572 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
IHC-Fr 1/200.
IHC-P 1/500.
WB 1/5000 - 1/10000. Detects a band of approximately 92 kDa (predicted molecular weight: 86 kDa).
ICC/IF 1/250.
IP 1/100.
  • Application notes
    Is unsuitable for Flow Cyt.
  • 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
    • Alternative names
      • 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

    Anti-beta Catenin antibody [E247] images



    • Predicted band size : 86 kDa

      Lane 1: Wild type HAP1 whole cell lysate (20 µg)
      Lane 2: CTNNB1 (β-catenin) knockout HAP1 whole cell lysate (20 µg)
      Lane 3: HeLa whole cell lysate (20 µg)
      Lane 4: A431 whole cell lysate (20 µg)

      Lanes 1 - 4: Merged signal (red and green). Green - ab32572 observed at 85 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab32572 was shown to specifically react with β-catenin when CTNNB1 (β-catenin) knockout samples were used. Wild-type and CTNNB1 (β-catenin) knockout samples were subjected to SDS-PAGE. ab32572 and ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at a 1/5000 dilution 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.

    • ab32572 staining in CTNNB1 (β-catenin) wild-type HAP1 cells (top panel) and in 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 ab32572 at 1/250 dilution and ab195889 at 1/250 dilution (shown in pseudocolour 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).

    • ab32572 staining beta-catenin in the bEnd.5 murine cell line by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.1% Triton in PBS and blocked with 10% serum for 30 minutes at 22°C. Samples were incubated with primary antibody (1/300) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.

      See Abreview

    • ab32572 staining beta Catenin in Dog colon tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 10% serum for 30 minutes at 25°C; antigen retrieval was by heat mediation in 10mM citrate buffer, pH6. Samples were incubated with primary antibody (1/250 in PBS with 1x casein) for 90 minutes at 25°C. A Biotin-conjugated Goat anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody.

      See Abreview



    • Performed under reducing conditions.

      Predicted band size : 86 kDa
      Observed band size : 90 kDa (why is the actual band size different from the predicted?)

      This image is courtesy of an anonymous Abreview

      Western blot image of ab32572 staining whole cell lysate of U2OS cells.  The gel was blocked with 5% milk for 1 hour at 21°C. The primary antibody was diluted 1/5000 and incubated  for 12 hours at 4°C.  A HRP conjugated swine anti-rabbit antibody was used as the secondary.

      See Abreview

    • ab32572 staining βcatenin in SW480 cells treated with BIO (ab120891), by ICC/IF. Increase of βcatenin expression correlates with increased concentration of BIO, as described in literature.
      The cells were incubated at 37°C for 48h in media containing different concentrations of ab120891 (BIO) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab32572 (1/200) dilution was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
    •     ab32572 at 1/200 staining mouse small intestine tissue sections by IHC-P. The tissue was formaldehyde fixed and a heat mediated antigen retrieval step was performed before incubation with the primary antibody. An HRP conjugated goat anti-rabbit antibody was used as the secondary.

      See Abreview

    • ab32572 staining beta Catenin in mouse liver tissue section by Immunohistochemistry (Frozen sections). Tissue samples were fixed with formaldehyde and blocked with 5% serum at 40C for 30 minutes. The sample was incubated with primary antibody (1/200) in dilution buffer containing PBS and 3% Goat Serum at 40C for 9 hours. An Alexa Fluor®488-conjugated Goat polyclonal to rabbit IgG was used as secondary antibody at 1/200 dilution.

      See Abreview

    • ab32572 staining human renal carcinoma tissue sections by IHC-P.  Sections were formaldehyde fixed and subjected to heat mediated antigen retrieval in citrate buffer (pH 6) prior to blocking with 1% milk for 45 minutes at 22°C.  The primary antibody was diluted 1/200 and incubated with the sample for 1 hour at 22°C.  A HRP conjugated goat anti-rabbit antibody, diluted 1/400, was used as the secondary.

      See Abreview

    • ab32572 staining β catenin in SKNSH cells treated with olanzapine (ab120736), by ICC/IF. Increase in expression of β catenin correlates with increased concentration of olanzapine, as described in literature.
      The cells were incubated at 37°C for 24h in media containing different concentrations of ab120736 (olanzapine) in DMSO, fixed with 4% formaldehyde for 10 minutes at room temperature and blocked with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 2h at room temperature. Staining of the treated cells with ab32572 (1/200 dilution) was performed overnight at 4°C in PBS containing 1% BSA and 0.1% tween. A DyLight 488 goat anti-rabbit polyclonal antibody (ab96899) at 1/250 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI and are shown in blue.
    • ab32572 showing positive staining in Cervical carcinoma tissue.

    • ab32572 showing positive staining in Breast carcinoma tissue.

    • ab32572 showing positive staining in Lung adenocarcinoma tissue.

    • ab32572 showing positive staining in Papillary carcinoma of thyroid gland tissue.

    • ab32572 showing positive staining in Kidney carcinoma tissue.

    References for Anti-beta Catenin antibody [E247] (ab32572)

    This product has been referenced in:
    • Huang H  et al. Tissue transglutaminase-1 promotes stemness and chemoresistance in gastric cancer cells by regulating Wnt/ß-catenin signaling. Exp Biol Med (Maywood) 242:194-202 (2017). Read more (PubMed: 27660242) »
    • Lu T  et al. Adamts18 deficiency promotes colon carcinogenesis by enhancing ß-catenin and p38MAPK/ERK1/2 signaling in the mouse model of AOM/DSS-induced colitis-associated colorectal cancer. Oncotarget 8:18979-18990 (2017). IHC ; Mouse . Read more (PubMed: 28145888) »

    See all 117 Publications for this product

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    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HeLa Cells)
    Loading amount
    20 µg
    Specification
    HeLa Cells
    Treatment
    10, 15, 20nM SiRNA Luciferase
    Gel Running Conditions
    Non-reduced Denaturing (7.5% gel)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Username

    Abcam user community

    Verified customer

    Submitted Feb 01 2012

    Thank you for contacting us.

    We recommend heat-mediated antigen retrieval: incubate sections in 10mM sodium citrate, pH 6,.0, for 20 minutes at 95-100oC before any blocking steps. For examples of staining achieved after heat-mediated antigen...

    Read More
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Huh7 cell line)
    Specification
    Huh7 cell line
    Fixative
    Paraformaldehyde
    Permeabilization
    Yes - 0.1% triton X-100 in PBS
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: RT°C
    Username

    Abcam user community

    Verified customer

    Submitted Jan 17 2012

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HepG2)
    Loading amount
    25 µg
    Specification
    HepG2
    Gel Running Conditions
    Reduced Denaturing (gel 10%)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted Jan 17 2012

    Thank you for contacting us. The concentration of lot GR39895-2 is XXXXX mg/ml. I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

    Application
    Western blot
    Sample
    Rhesus monkey Tissue lysate - whole (Brain Tissue - Isolated striatum)
    Loading amount
    20 µg
    Specification
    Brain Tissue - Isolated striatum
    Gel Running Conditions
    Reduced Denaturing (7% Tris Acetate Gel)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted Jan 21 2011

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