遺伝子ノックアウト細胞株 検証済

Anti-beta Catenin 抗体 [15B8] (ab6301)

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Key features and details

  • Mouse monoclonal [15B8] to beta Catenin
  • Suitable for: WB, IHC-P, EMSA
  • Knockout validated
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG1

製品の概要

  • 製品名

    Anti-beta Catenin antibody [15B8]
    beta Catenin 一次抗体 製品一覧

  • 製品の詳細

    Mouse monoclonal [15B8] to beta Catenin

  • 由来種

    Mouse

  • アプリケーション

    適用あり: WB, IHC-P, EMSAmore details

  • 種交差性

    交差種: Mouse, Rat, Human
    交差が予測される動物種: Cow, Dog, Zebrafish

  • 免疫原

    Recombinant full length protein corresponding to Chicken beta Catenin.

  • ポジティブ･コントロール

    • WB: Hap1, HeLa, A43, HEK293, Caco2, NIH3T3 and PC12 cell lysates.

  • 特記事項

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab6301 in the following tested applications.

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

アプリケーション Abreviews 特記事項
WB Use a concentration of 1 µg/ml. Detects a band of approximately 95 kDa (predicted molecular weight: 85 kDa).
IHC-P Use at an assay dependent concentration. PubMed: 21205793
EMSA Use at an assay dependent concentration. PubMed: 12533679

ターゲット情報

  • 機能

    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.

  • 組織特異性

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

  • 関連疾患

    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.

  • 配列類似性

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

  • 翻訳後修飾

    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.

  • 細胞内局在

    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

  • 参照データベース

    see all

  • 別名

    • 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

画像

  • Western blot - Anti-beta Catenin antibody [15B8] (ab6301)
    Western blot - Anti-beta Catenin antibody [15B8] (ab6301)
    All lanes : Anti-beta Catenin antibody [15B8] (ab6301) at 1 µg/ml

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : CTNNB1 knockout HAP1 cell lysate
    Lane 3 : Wild-type HeLa cell lysate
    Lane 4 : CTNNB1 knockout HeLa cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 85 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab6301 observed at 95 kDa. Red - loading control, ab181602 observed at 37 kDa.

     ab6301 was shown to react with beta Catenin in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255352 (knockout cell lysate ab263756) was used. Wild-type and beta Catenin knockout samples were subjected to SDS-PAGE. ab6301 and Anti-GAPDH antibody EPR16891] - Loading Control (ab181602) were incubated overnight at 4°C at 1 µg/ml and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

     

     

  • Western blot - Anti-beta Catenin antibody [15B8] (ab6301)
    Western blot - Anti-beta Catenin antibody [15B8] (ab6301)
    All lanes : Anti-beta Catenin antibody [15B8] (ab6301) at 1 µg/ml

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : CTNNB1 knockout HAP1 whole cell lysate
    Lane 3 : HeLa whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 85 kDa



    Lanes 1 - 3: Merged signal (red and green). Green - ab6301 observed at 85 kDa. Red - loading control, ab181602, observed at 37 kDa.

    ab6301 was shown to specifically react with beta-Catenin in wild-type HAP1 cells along with additional cross reactive bands. No bands were observed when knockout samples were used. Wild-type and beta Catenin knockout samples were subjected to SDS-PAGE. Ab6301 and ab181602 (Rabbit 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-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-beta Catenin antibody [15B8] (ab6301)
    Western blot - Anti-beta Catenin antibody [15B8] (ab6301)
    All lanes : Anti-beta Catenin antibody [15B8] (ab6301) at 1 µg/ml

    Lane 1 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 3 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate
    Lane 4 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 5 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal Secondary Antibody to Mouse IgG - H&L (HRP), pre-adsorbed at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 85 kDa
    Observed band size: 95 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 73 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 4 minutes


    This blot was produced using a 10% 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 5% Bovine Serum Albumin before being incubated with ab6301 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

参考文献

ab6301 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab6301 は 29 報の論文で使用されています。

  • Zhang C  et al. HEF1 regulates differentiation through the Wnt5a/ß-catenin signaling pathway in human gastric cancer. Biochem Biophys Res Commun 509:201-208 (2019). PubMed: 30579603
  • Song J  et al. Long noncoding RNA GAS5 inhibits angiogenesis and metastasis of colorectal cancer through the Wnt/ß-catenin signaling pathway. J Cell Biochem N/A:N/A (2019). PubMed: 30672001
  • Sun K  et al. Tre2 (USP6NL) promotes colorectal cancer cell proliferation via Wnt/ß-catenin pathway. Cancer Cell Int 19:102 (2019). PubMed: 31015802
  • Chen J  et al. IL-6/YAP1/ß-catenin signaling is involved in intervertebral disc degeneration. J Cell Physiol 234:5964-5971 (2019). PubMed: 30511395
  • Li Y  et al. Discs large 1 controls daughter-cell polarity after cytokinesis in vertebrate morphogenesis. Proc Natl Acad Sci U S A 115:E10859-E10868 (2018). PubMed: 30377270
View all Publications for this product

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1-4 of 4 Abreviews or Q&A

Immunohistochemistry (Frozen sections) abreview for Anti-beta Catenin antibody [15B8]

 Excellent
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (Heart)
Permeabilization
Yes - Triton 0.1%
Specification
Heart
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Paraformaldehyde
Read More

Abcam user community

Verified customer

投稿 Dec 07 2017

Western blot abreview for Anti-beta Catenin antibody [15B8]

 Excellent
Abreviews
abreview image
Application
Western blot
Sample
Cow Tissue lysate - other (Liver membrane)
Gel Running Conditions
Reduced Denaturing (12)
Loading amount
15 µg
Specification
Liver membrane
Blocking step
Milk as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 1.5% · Temperature: 25°C
Read More

Abcam user community

Verified customer

投稿 Nov 22 2017

Immunohistochemistry (Frozen sections) abreview for Anti-beta Catenin antibody [15B8]

 Excellent
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (foreskin)
Permeabilization
No
Specification
foreskin
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 22°C
Fixative
none
Read More

Mr. Julien Coutier

Verified customer

投稿 Jul 05 2017

Immunohistochemistry (Frozen sections) abreview for Anti-beta Catenin antibody [15B8]

 Poor
Abreviews
abreview image
Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (skin)
Permeabilization
Yes - PBST
Specification
skin
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Acetone
Read More

Abcam user community

Verified customer

投稿 Jan 20 2017

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