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遺伝子ノックアウト細胞株 検証済リコンビナントRabMAb

Anti-CBL 抗体 [YE323] - C-terminal (ab32027)

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Western blot - Anti-CBL antibody [YE323] - C-terminal (ab32027)
  • Western blot - Anti-CBL antibody [YE323] - C-terminal (ab32027)
  • Western blot - Anti-CBL antibody [YE323] - C-terminal (ab32027)
  • Immunocytochemistry/ Immunofluorescence - Anti-CBL antibody [YE323] - C-terminal (ab32027)
  • Flow Cytometry (Intracellular) - Anti-CBL antibody [YE323] - C-terminal (ab32027)
  • Western blot - Anti-CBL antibody [YE323] - C-terminal (ab32027)
  • Western blot - Anti-CBL antibody [YE323] - C-terminal (ab32027)
  • Anti-CBL antibody [YE323] - C-terminal (ab32027)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [YE323] to CBL - C-terminal
  • Suitable for: WB, Flow Cyt (Intra), ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 647 Carrier Free PE

製品の概要

  • 製品名

    Anti-CBL antibody [YE323] - C-terminal
    CBL 一次抗体 製品一覧

  • 製品の詳細

    Rabbit monoclonal [YE323] to CBL - C-terminal

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: WB, Flow Cyt (Intra), ICC/IFmore details
    適用なし: IP

  • 種交差性

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

  • 免疫原

    Synthetic peptide within Human CBL aa 850 to the C-terminus (C terminal). The exact sequence is proprietary.
    Database link: P22681

  • エピトープ

    ab32027 reacts with an epitope located in the C terminal region of CBL.

  • ポジティブ･コントロール

    • WB: HEK293T, HAP1, Jurkat, THP-1, WEHI-231, F9 and Raji cell lysates; Mouse thymus tissue lysate, Rat testis lysate, Rat thymus lysate. ICC/IF: Jurkat cells. Flow Cyt (intra): Jurkat cells.

  • 特記事項

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

製品の特性

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab32027の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
WB
1/1000. Detects a band of approximately 120 kDa (predicted molecular weight: 99 kDa).

For unpurified use at 1/5000
Flow Cyt (Intra)
1/30.

For unpurified use at 1/100  ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ICC/IF
1/50.

For unpurified use at 1/100
特記事項
WB
1/1000. Detects a band of approximately 120 kDa (predicted molecular weight: 99 kDa).

For unpurified use at 1/5000
Flow Cyt (Intra)
1/30.

For unpurified use at 1/100  ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ICC/IF
1/50.

For unpurified use at 1/100
追加情報
Is unsuitable for IP.

ターゲット情報

  • 機能

    Participates in signal transduction in hematopoietic cells. Adapter protein that functions as a negative regulator of many signaling pathways that start from receptors at the cell surface. Acts as an E3 ubiquitin-protein ligase, which accepts ubiquitin from specific E2 ubiquitin-conjugating enzymes, and then transfers it to substrates promoting their degradation by the proteasome. Recognizes activated receptor tyrosine kinases, including PDGFA, EGF and CSF1, and terminates signaling.

  • パスウェイ

    Protein modification; protein ubiquitination.

  • 関連疾患

    Defects in CBL are the cause of Noonan syndrome-like disorder (NSL) [MIM:613563]. NSL is a syndrome characterized by a phenotype reminiscent of Noonan syndrome. Clinical features are highly variable, including facial dysmorphism, short neck, developmental delay, hyperextensible joints and thorax abnormalities with widely spaced nipples. The facial features consist of triangular face with hypertelorism, large low-set ears, ptosis, and flat nasal bridge. Some patients manifest cardiac defects.

  • 配列類似性

    Contains 1 Cbl-PTB (Cbl-type phosphotyrosine-binding) domain.
    Contains 1 RING-type zinc finger.
    Contains 1 UBA domain.

  • ドメイン

    The RING-type zinc finger domain mediates binding to an E2 ubiquitin-conjugating enzyme.
    The N-terminus is composed of the phosphotyrosine binding (PTB) domain, a short linker region and the RING-type zinc finger. The PTB domain, which is also called TKB (tyrosine kinase binding) domain, is composed of three different subdomains: a four-helix bundle (4H), a calcium-binding EF hand and a divergent SH2 domain.

  • 翻訳後修飾

    Phosphorylated on tyrosine residues by EGFR, SYK, FYN and ZAP70 (By similarity). Phosphorylated on tyrosine residues by INSR.

  • 細胞内局在

    Cytoplasm.
  • Information by UniProt

  • 参照データベース

    see all

  • 別名

    • 4732447J05Rik antibody
    • C CBL antibody
    • Cas Br M (murine) ecotropic retroviral transforming sequence antibody
    • Casitas B lineage lymphoma proto oncogene antibody
    • Casitas B-lineage lymphoma proto-oncogene antibody
    • CBL 2 antibody
    • cbl antibody
    • CBL_HUMAN antibody
    • CBL2 antibody
    • E3 ubiquitin protein ligase CBL antibody
    • E3 ubiquitin-protein ligase CBL antibody
    • Oncogene CBL2 antibody
    • Proto oncogene c CBL antibody
    • Proto-oncogene c-CBL antibody
    • RGD1561386 antibody
    • RING finger protein 55 antibody
    • RNF55 antibody
    • Signal transduction protein CBL antibody
    see all

画像

  • Western blot - Anti-CBL antibody [YE323] - C-terminal (ab32027)
    Western blot - Anti-CBL antibody [YE323] - C-terminal (ab32027)
    All lanes : Anti-CBL antibody [YE323] - C-terminal (ab32027) at 1/1000 dilution

    Lane 1 : Wild-type HCT 116 cell lysate
    Lane 2 : CBL knockout HCT 116 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 99 kDa
    Observed band size: 105 kDa why is the actual band size different from the predicted?



    Western blot: Anti-CBL antibody [YE323] (ab32027) staining at 1/1000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab32027 was shown to bind specifically to CBL. A band was observed at 105 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in CBL knockout cell line. To generate this image, wild-type and CBL knockout 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

  • Western blot - Anti-CBL antibody [YE323] - C-terminal (ab32027)
    Western blot - Anti-CBL antibody [YE323] - C-terminal (ab32027)
    All lanes : Anti-CBL antibody [YE323] - C-terminal (ab32027) at 1/1000 dilution

    Lane 1 : Raji (Human Burkitt's lymphoma B lymphocyte) whole cell lysate at 15 µg
    Lane 2 : Rat testis lysate at 20 µg
    Lane 3 : Rat thymus lysate at 20 µg

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/2000 dilution

    Predicted band size: 99 kDa
    Observed band size: 110 kDa why is the actual band size different from the predicted?

  • Western blot - Anti-CBL antibody [YE323] - C-terminal (ab32027)
    Western blot - Anti-CBL antibody [YE323] - C-terminal (ab32027)
    All lanes : Anti-CBL antibody [YE323] - C-terminal (ab32027) at 1/1000 dilution (unpurified)

    Lane 1 : Wild-type HEK293T cell lysate
    Lane 2 : CBL knockout HEK293T cell lysate
    Lane 3 : THP-1 cell lysate
    Lane 4 : Raji cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 99 kDa
    Observed band size: 110 kDa why is the actual band size different from the predicted?



    Lanes 1-4: Merged signal (red and green). Green - ab32027 observed at 110 kDa. Red - loading control ab8245 observed at 36 kDa.

    Unpurified ab32027 Anti-CBL antibody [YE323] - C-terminal was shown to specifically react with CBL in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab267245 (knockout cell lysate ab257200) was used. Wild-type and CBL knockout samples were subjected to SDS-PAGE. ab32027 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution 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.

  • Immunocytochemistry/ Immunofluorescence - Anti-CBL antibody [YE323] - C-terminal (ab32027)
    Immunocytochemistry/ Immunofluorescence - Anti-CBL antibody [YE323] - C-terminal (ab32027)
    Immunocytochemistry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling CBL with purified ab32027 at 1/50 dilution (4.26 µg/mL). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 µg/mL). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/mL) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
  • Flow Cytometry (Intracellular) - Anti-CBL antibody [YE323] - C-terminal (ab32027)
    Flow Cytometry (Intracellular) - Anti-CBL antibody [YE323] - C-terminal (ab32027)

    Intracellular Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) cells labeling CBL with purified ab32027 at 1/30 dilution (10 µg/mL) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

     

  • Western blot - Anti-CBL antibody [YE323] - C-terminal (ab32027)
    Western blot - Anti-CBL antibody [YE323] - C-terminal (ab32027)

    Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: CBL knockout HAP1 whole cell lysate (20 µg)
    Lane 3: THP1 whole cell lysate (20 µg)
    Lane 4: Raji whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab32027 observed at 100 kDa. Red - loading control, ab9484, observed at 37 kDa.

    Unpurified ab32027 was shown to specifically react with CBL in wild-type HAP1 cells as signal was lost in CBL knockout cells. Wild-type and CBL knockout samples were subjected to SDS-PAGE. ab32027 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 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.

  • Western blot - Anti-CBL antibody [YE323] - C-terminal (ab32027)
    Western blot - Anti-CBL antibody [YE323] - C-terminal (ab32027)
    All lanes : Anti-CBL antibody [YE323] - C-terminal (ab32027) at 1/1000 dilution

    Lane 1 : WEHI-231 (Mouse B cell lymphoma B lymphocyte) cell lysate
    Lane 2 : F9 (Mouse embryonal carcinoma epithelial cell) cell lysate
    Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) cell lysate
    Lane 4 : Mouse thymus lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 99 kDa
    Observed band size: 110 kDa why is the actual band size different from the predicted?



    Blocking and diluting buffer and concentration: 5% NFDM/TBST

    Exposure time: 30 seconds

  • Anti-CBL antibody [YE323] - C-terminal (ab32027)
    Anti-CBL antibody [YE323] - C-terminal (ab32027)

プロトコール

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

参考文献 (13)

ab32027 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。

ab32027 は 13 報の論文で使用されています。

  • Nie XH  et al. Paeoniflorin Regulates NEDD4L/STAT3 Pathway to Induce Ferroptosis in Human Glioma Cells. J Oncol 2022:6093216 (2022). PubMed: 36618071
  • Jin Q  et al. Overexpression of E3 ubiquitin ligase Cbl attenuates endothelial dysfunction in diabetes mellitus by inhibiting the JAK2/STAT4 signaling and Runx3-mediated H3K4me3. J Transl Med 19:469 (2021). PubMed: 34798872
  • Lei X  et al. circTLK1 facilitates the proliferation and metastasis of renal cell carcinoma by regulating miR-495-3p/CBL axis. Open Life Sci 16:362-374 (2021). PubMed: 33954256
  • Xie G  et al. Conjugated secondary 12a-hydroxylated bile acids promote liver fibrogenesis. EBioMedicine 66:103290 (2021). PubMed: 33752128
  • Zhang D  et al. MicroRNA-935 Directly Targets FZD6 to Inhibit the Proliferation of Human Glioblastoma and Correlate to Glioma Malignancy and Prognosis. Front Oncol 11:566492 (2021). PubMed: 33791198
View all Publications for this product

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