Anti-beta Catenin (phospho Y654) antibody [1B11] (ab24925)

Overview

  • Product nameAnti-beta Catenin (phospho Y654) antibody [1B11]See all beta Catenin primary antibodies ...
  • Description
    Mouse monoclonal [1B11] to beta Catenin (phospho Y654)
  • Specificityab24925 recognises beta catenin phosphorylated at tyrosine 654.
  • Tested applicationsIHC-P, ICC/IF, IHC-Fr, WB, ELISA, ICC more details
  • Species reactivity
    Reacts with: Mouse, Rat, Dog, Human
  • Immunogen

    Synthetic phosphopeptide

  • Epitopeab24925 reacts with an epitope located in the region of phosphorylated tyrosine 654.
  • Positive control
    • Vanadate treated SW480 cell lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab24925 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Notes
IHC-P Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration. PubMed: 21970630
IHC-Fr Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Predicted molecular weight: 86 kDa.
ELISA Use a concentration of 0.05 µg/ml.
ICC 1/50.

Target

  • FunctionKey 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 specificityExpressed 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 diseaseDefects 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 similaritiesBelongs 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 localizationCytoplasm. 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
  • 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

Anti-beta Catenin (phospho Y654) antibody [1B11] images

  • Immunohistochemical analysis of rat bladder tissue sections, staining beta Catenin (phospho Y654) with ab24925.

    Tissue was fixed with 10% Neutral Buffered Formalin and blocked with 5000 µg/ml BSA for 30 minutes at 22°C. Samples were incubated with primary antibody (1/100 in diluent) for 1 hour at 22°C. An AlexaFluor®488-conjugated goat anti-mouse polyclonal IgG (1/400) was used as the secondary antibody.

    See Abreview

  • All lanes : Anti-beta Catenin (phospho Y654) antibody [1B11] (ab24925) at 0.5 µg/ml

    Lane 1 : Whole cell extracts of control OVCAR 5 tumor cells
    Lane 2 : Whole cell extracts pervanadate treated OVCAR 5 tumor cells

    Lysates/proteins at 20000 cells per lane.

    Secondary
    A HRP-conjugated antibody
    developed using the ECL technique

    Predicted band size : 86 kDa


    Exposure time : 30 seconds
    Whole cell extracts of control (1) or pervanadate treated (2) OVCAR 5 tumor cells were applied to SDS-PAGE (ca 20,000 cells per lane) and transferred to a PVDF membrane. The immunoblot was probed with ab24925 (0.5ug/ml) for 1 hour at room temp. and developed by ECL (exp. time: 30 seconds). The blot was probed for 1 hour at RT.

References for Anti-beta Catenin (phospho Y654) antibody [1B11] (ab24925)

This product has been referenced in:
  • Chen R  et al. Ubiquitin-mediated interaction of p210 BCR/ABL with ß-catenin supports disease progression in a murine model for chronic myelogenous leukemia. Blood 122:2114-24 (2013). Read more (PubMed: 23950177) »
  • Brunet T  et al. Evolutionary conservation of early mesoderm specification by mechanotransduction in Bilateria. Nat Commun 4:2821 (2013). IHC (PFA fixed) ; Fruit fly (Drosophila melanogaster) . Read more (PubMed: 24281726) »

See all 10 Publications for this product

Product Wall


We usually add a Ready-to-use cocktails of inhibitors which will contain Orthovanadate, or you can prepare it yourself :

Na Orthovanadate in 1mM concentration.

Sodium orthovanadate preparation

All steps to be performed i...

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Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Rat Tissue sections (Bladder)
Specification Bladder
Fixative 10% Buffered Formalin
Antigen retrieval step None
Permeabilization No
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 5000µg/mL · Temperature: 22°C
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I am very pleased to hear you would like to accept our offer and test ab24925 in IHC of paraffin-embedded tissue (IHC-P). To redeem this offer, please submit an Abreview for IHC...

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Application Western blot
Sample Human Cell lysate - whole cell (caco-2)
Loading amount 50 µg
Specification caco-2
Gel Running Conditions Reduced Denaturing (8%)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 27°C
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Submitted Sep 21 2012

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I'm glad to hear that this antibody is working for Western blot. I would recommend going ahead and using the stored antibody, as one additi...

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