使用敲除细胞株进行验证

Anti-beta Catenin抗体[12F7] (ab22656)

概述

性能

应用

Our Abpromise guarantee covers the use of ab22656 in the following tested applications.

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

应用 Ab评论 说明
Flow Cyt Use 1µg for 106 cells. ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.
ICC/IF Use at an assay dependent concentration. PubMed: 24158438
IHC-P Use a concentration of 1 - 5 µg/ml. Perform heat mediated antigen retrieval via the microwave method before commencing with IHC staining protocol.
WB Use a concentration of 2 µg/ml. Detects a band of approximately 85 kDa (predicted molecular weight: 85 kDa).

We recommend using 3% milk as the blocking agent for Western blot.

IP Use at an assay dependent concentration. Use at a concentration of 3.3 - 5 µg/ml cell extract.

靶标

  • 功能
    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.
  • 组织特异性
    Expressed in several hair follicle cell types: basal and peripheral matrix cells, and cells of the outer and inner root sheaths. Expressed in colon.
  • 疾病相关
    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.
  • 序列相似性
    Belongs to the beta-catenin family.
    Contains 12 ARM repeats.
  • 翻译后修饰
    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.
  • 细胞定位
    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
  • 数据库链接
  • 别名
    • 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 [12F7] 图像



  • Predicted band size : 85 kDa

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: CTNNB1 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 - ab22656 observed at 95 kDa. Red - loading control, ab181602, observed at 37 kDa.

    ab22656 was shown to recognize CTNNB1 when CTNNB1 knockout samples were used, along with additional cross-reactive bands. Wild-type and CTNNB1 knockout samples were subjected to SDS-PAGE. Ab22656 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/10000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • All lanes : Anti-beta Catenin antibody [12F7] (ab22656) at 2 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : DU 145 (Human prostate carcinoma cell line) Whole Cell Lysate
    Lane 3 : A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 85 kDa
    Observed band size : 85 kDa


    Exposure time : 12 minutes

    We recommend using 3% milk as the blocking agent for Western blot.

  • IHC image of ab22656 staining beta Catenin in human colon adenocarcinoma formalin-fixed paraffin-embedded tissue sections*, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab22656, 1/500 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. High magnification of the tumor region - T (lower right panel) and adjacent normal crypts - N (lower left panel) are shown.
    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

  • Overlay histogram showing HeLa cells stained with ab22656 (red line). The cells were fixed with methanol (5 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 (ab22656, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • IHC image of Ab22656 staining in human normal colon formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab22656, 10µg/ml, 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.

    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.
  • ab22656 staining rat bowel tissue section by IHC-P.  The section was formaldehyde fixed and subjected to heat mediated antigen retrieval in citrate buffer (pH6) prior to blocking with 10% serum for 10 hours at 24°C.  The primary antibody was diluted 1/500 in milk and incubated with the section for 12 hours at 4°C.  A biotinylated rabbit anti-mouse was used as the secondart antibody.

    See Abreview

Anti-beta Catenin antibody [12F7] (ab22656)参考文献

This product has been referenced in:
  • Meng X  et al. Heterogeneous nuclear ribonucleoprotein A1 interacts with microRNA-34a to promote chondrogenic differentiation of mesenchymal stem cells. Am J Transl Res 9:1774-1782 (2017). WB ; Mouse . Read more (PubMed: 28469782) »
  • Feng T  et al. Decrease in stathmin expression by arsenic trioxide inhibits the proliferation and invasion of osteosarcoma cells via the MAPK signal pathway. Oncol Lett 14:1333-1340 (2017). Read more (PubMed: 28789348) »

See all 18 Publications for this product

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Thank you for contacting us. Having checked our catalog for the appropriate products, I would like to send you the following recommendations. Where possible I looked for both monoclonal and polyclonal antibodies, so if not mentioned we unfortunately d...

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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 (Kidney cancer cell)
Loading amount
20 µg
Specification
Kidney cancer cell
Gel Running Conditions
Non-reduced Denaturing (10% 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

提交于 Nov 11 2010

Application
Western blot
Sample
Mouse Cell lysate - nuclear (MEF)
Loading amount
100 µg
Specification
MEF
Gel Running Conditions
Non-reduced Denaturing (gel 10%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

提交于 Mar 06 2008

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (bowel)
Specification
bowel
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrat puffer ph 6
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Username

Abcam user community

Verified customer

提交于 Feb 05 2008

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rat Tissue sections (bowel)
Specification
bowel
Fixative
Formaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer pH 6
Permeabilization
No
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Username

Abcam user community

Verified customer

提交于 Jan 31 2008

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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