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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-NCE2/UBE2F 抗体 [EPR12932] (ab185234)

Submit a review Submit a question References (5)
Western blot - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)
  • Flow Cytometry (Intracellular) - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)
  • Immunocytochemistry/ Immunofluorescence - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)
  • Western blot - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)
  • Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR12932] to NCE2/UBE2F
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

製品の概要

  • 製品名

    Anti-NCE2/UBE2F antibody [EPR12932]
    NCE2/UBE2F 一次抗体 製品一覧

  • 製品の詳細

    Rabbit monoclonal [EPR12932] to NCE2/UBE2F

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details

  • 種交差性

    交差種: Human

  • 免疫原

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • ポジティブ･コントロール

    • WB: HeLa, Jurkat, A375 and NTERA2/D1 cell lysates. Flow Cyt (intra): A375 cells. ICC/IF: A375 cells. IHC-P: Human brain and Human renal carcinoma tissues.

  • 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ab185234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
Flow Cyt (Intra)
1/160.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB
1/1000. Detects a band of approximately 21 kDa (predicted molecular weight: 21 kDa).
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/250.
特記事項
Flow Cyt (Intra)
1/160.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB
1/1000. Detects a band of approximately 21 kDa (predicted molecular weight: 21 kDa).
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF
1/250.

ターゲット情報

  • 機能

    Accepts the ubiquitin-like protein NEDD8 from the UBA3-NAE1 E1 complex and catalyzes its covalent attachment to other proteins. The specific interaction with the E3 ubiquitin ligase RBX2, but not RBX1, suggests that the RBX2-UBE2F complex neddylates specific target proteins, such as CUL5.

  • 組織特異性

    Widely expressed (at protein level).

  • パスウェイ

    Protein modification; protein neddylation.

  • 配列類似性

    Belongs to the ubiquitin-conjugating enzyme family. UBE2F subfamily.
  • Information by UniProt

  • 参照データベース

    • 別名

      • NCE2 antibody
      • NEDD8 carrier protein UBE2F antibody
      • NEDD8 conjugating enzyme antibody
      • NEDD8 conjugating enzyme UBE2F antibody
      • NEDD8 protein ligase UBE2F antibody
      • NEDD8-conjugating enzyme 2 antibody
      • NEDD8-conjugating enzyme UBE2F antibody
      • ube2f antibody
      • UBE2F_HUMAN antibody
      • Ubiquitin conjugating enzyme E2F antibody
      • Ubiquitin conjugating enzyme E2F (putative) antibody
      • Ubiquitin-conjugating enzyme E2 F antibody
      see all

    画像

    • Western blot - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)
      Western blot - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)
      All lanes : Anti-NCE2/UBE2F antibody [EPR12932] (ab185234) at 1/1000 dilution

      Lane 1 : Wild-type HeLa cell lysate
      Lane 2 : UBE2F knockout HeLa cell lysate
      Lane 3 : Jurkat 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: 21 kDa
      Observed band size: 24 kDa why is the actual band size different from the predicted?



      Lanes 1-3: Merged signal (red and green). Green - ab185234 observed at 24 kDa. Red - loading control ab8245 observed at 36 kDa. 

       ab185234 Anti-NCE2/UBE2F antibody [EPR12932] was shown to specifically react with NCE2/UBE2F in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265339 (knockout cell lysate ab257777) was used.  Wild-type and NCE2/UBE2F knockout samples were subjected to SDS-PAGE.  ab185234 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.

    • Flow Cytometry (Intracellular) - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)
      Flow Cytometry (Intracellular) - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)

      Intracellular flow cytometric analysis of A375 cells (2% paraformaldehyde-fixed) labeling NCE2/UBE2F with ab185234at 1/160 dilution (red) or a rabbit IgG (negative) (green), followed by Goat anti rabbit IgG (FITC) secondary at 1/150 dilution. 

       

       

    • Immunocytochemistry/ Immunofluorescence - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)
      Immunocytochemistry/ Immunofluorescence - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)

      Immunofluorescent analysis of A375 cells (4% paraformaldehyde-fixed) labeling NCE2/UBE2F with ab185234 at 1/250 dilution followed by Goat anti rabbit IgG (AlexaFluor® 555) secondary at 1/200 dilution and counter-stained with DAPI (blue).

    • Western blot - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)
      Western blot - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)
      All lanes : Anti-NCE2/UBE2F antibody [EPR12932] (ab185234) at 1/1000 dilution

      Lane 1 : Jurkat cell lysate
      Lane 2 : A375 cell lysate
      Lane 3 : NTERA2/D1 cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

      Predicted band size: 21 kDa

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)

      Immunohistochemical analysis of paraffin-embedded Human renal carcinoma labeling NCE2/UBE2F with ab185234 at 1/100 dilution followed by pre-diluted HRP-conjugated secondary antibody and counter-stained with Hematoxylin.

      Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)

      Immunohistochemical analysis of paraffin-embedded Human brain labeling NCE2/UBE2F with ab185234 at 1/100 dilution followed by pre-diluted HRP-conjugated secondary antibody and counter-stained with Hematoxylin.

      Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

    • Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)
      Anti-NCE2/UBE2F antibody [EPR12932] (ab185234)

    参考文献 (5)

    ab185234 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。

    ab185234 は 5 報の論文で使用されています。

    • Varland S  et al. N-terminal acetylation shields proteins from degradation and promotes age-dependent motility and longevity. Nat Commun 14:6774 (2023). PubMed: 37891180
    • Guo Z  et al. HUWE1-dependent DNA-PKcs neddylation modulates its autophosphorylation in DNA damage response. Cell Death Dis 11:400 (2020). PubMed: 32457294
    • Zhou L  et al. Induction of NEDD8-conjugating enzyme E2 UBE2F by platinum protects lung cancer cells from apoptosis and confers to platinum-insensitivity. Cell Death Dis 11:975 (2020). PubMed: 33184273
    • Kabir S  et al. The CUL5 ubiquitin ligase complex mediates resistance to CDK9 and MCL1 inhibitors in lung cancer cells. Elife 8:N/A (2019). PubMed: 31294695
    • Zhou W  et al. UBE2M Is a Stress-Inducible Dual E2 for Neddylation and Ubiquitylation that Promotes Targeted Degradation of UBE2F. Mol Cell 70:1008-1024.e6 (2018). PubMed: 29932898

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