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リコンビナントRabMAb

Anti-E2F6 抗体 [EPR25340-17] (ab289963)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E2F6 antibody [EPR25340-17] (ab289963)
  • Immunocytochemistry/ Immunofluorescence - Anti-E2F6 antibody [EPR25340-17] (ab289963)
  • Flow Cytometry (Intracellular) - Anti-E2F6 antibody [EPR25340-17] (ab289963)
  • Immunoprecipitation - Anti-E2F6 antibody [EPR25340-17] (ab289963)
  • Western blot - Anti-E2F6 antibody [EPR25340-17] (ab289963)
  • Flow Cytometry (Intracellular) - Anti-E2F6 antibody [EPR25340-17] (ab289963)
  • Immunocytochemistry/ Immunofluorescence - Anti-E2F6 antibody [EPR25340-17] (ab289963)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E2F6 antibody [EPR25340-17] (ab289963)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E2F6 antibody [EPR25340-17] (ab289963)
  • Immunoprecipitation - Anti-E2F6 antibody [EPR25340-17] (ab289963)
  • Western blot - Anti-E2F6 antibody [EPR25340-17] (ab289963)
  • Anti-E2F6 antibody [EPR25340-17] (ab289963)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR25340-17] to E2F6
  • Suitable for: Flow Cyt (Intra), ICC/IF, WB, IHC-P, IP
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Carrier Free

製品の概要

  • 製品名

    Anti-E2F6 antibody [EPR25340-17]
    E2F6 一次抗体 製品一覧

  • 製品の詳細

    Rabbit monoclonal [EPR25340-17] to E2F6

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: Flow Cyt (Intra), ICC/IF, WB, IHC-P, IPmore details
    適用なし: ChIP

  • 種交差性

    交差種: Mouse, Rat, Human

  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • ポジティブ･コントロール

    • WB: MEF, NIH/3T3, C2C12, RAW264.7, PC-12, Jurkat ,K562, HeLa and HepG2 whole cell lysates; Mouse E17 brain tissue lysate. IHC-P: Human cerebrum tissue; Mouse cerebrum tissue; Rat cerebrum tissue. Flow cyt: Jurkat and NIH/3T3 cells. ICC/IF: Jurkat and NIH/3T3 cells. IP: Jurkat and NIH/3T3 whole cell lysates.

  • 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ab289963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)
1/500.
ICC/IF
1/100.
WB
1/1000. Detects a band of approximately 35 kDa (predicted molecular weight: 32 kDa).
IHC-P
1/5000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
1/30.
特記事項
Flow Cyt (Intra)
1/500.
ICC/IF
1/100.
WB
1/1000. Detects a band of approximately 35 kDa (predicted molecular weight: 32 kDa).
IHC-P
1/5000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP
1/30.
追加情報
Is unsuitable for ChIP.

ターゲット情報

  • 機能

    Inhibitor of E2F-dependent transcription. Binds DNA cooperatively with DP proteins through the E2 recognition site, 5'-TTTC[CG]CGC-3'. Has a preference for the 5'-TTTCCCGC-3' E2F recognition site. E2F-6 lacks the transcriptional activation and pocket protein binding domains. Appears to regulate a subset of E2F-dependent genes whose products are required for entry into the cell cycle but not for normal cell cycle progression. May silence expression via the recruitment of a chromatin remodeling complex containing histone H3-K9 methyltransferase activity. Overexpression delays the exit of cells from the S-phase.

  • 組織特異性

    Expressed in all tissues examined. Highest levels in placenta, skeletal muscle, heart, ovary, kidney, small intestine and spleen.

  • 配列類似性

    Belongs to the E2F/DP family.

  • 細胞内局在

    Nucleus.
  • Information by UniProt

  • 参照データベース

    • 別名

      • E2F 6 antibody
      • E2F binding site modulating activity protein antibody
      • E2F transcription factor 6 antibody
      • E2F transcription factor 6 isoform 1 antibody
      • E2F-6 antibody
      • E2F6 antibody
      • E2F6_HUMAN antibody
      • EMA antibody
      • MGC111545 antibody
      • Transcription factor E2F6 antibody
      see all

    画像

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E2F6 antibody [EPR25340-17] (ab289963)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E2F6 antibody [EPR25340-17] (ab289963)

      Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labelling E2F6 with ab289963 at 1/5000dilution, followed by LeicaDS9800 (Bond®, Polymer Refine Detection). Nuclear staining on rat cerebrum. The section was incubated with ab289963 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is LeicaDS9800 (Bond®, Polymer Refine Detection).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 10 mins.

    • Immunocytochemistry/ Immunofluorescence - Anti-E2F6 antibody [EPR25340-17] (ab289963)
      Immunocytochemistry/ Immunofluorescence - Anti-E2F6 antibody [EPR25340-17] (ab289963)

      Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 cells labeling E2F6 with ab289963 at 1/100 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in NIH/3T3 cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

      Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

    • Flow Cytometry (Intracellular) - Anti-E2F6 antibody [EPR25340-17] (ab289963)
      Flow Cytometry (Intracellular) - Anti-E2F6 antibody [EPR25340-17] (ab289963)

      Intracellular flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized NIH/3T3 (Mouse embryonic fibroblast) cells labelling E2F6 with ab289963 at 1/500 dilution (0.1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

    • Immunoprecipitation - Anti-E2F6 antibody [EPR25340-17] (ab289963)
      Immunoprecipitation - Anti-E2F6 antibody [EPR25340-17] (ab289963)

      E2F6 was immunoprecipitated from Jurkat (human T cell leukemia T lymphocyte) whole cell lysate with ab289963 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab289963 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

      Lane 1: Jurkat (human T cell leukemia T lymphocyte) whole cell lysate 10 µg

      Lane 2: ab289963 IP in Jurkat whole cell lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab289963 in Jurkat whole cell lysate

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 3.25 seconds

       

    • Western blot - Anti-E2F6 antibody [EPR25340-17] (ab289963)
      Western blot - Anti-E2F6 antibody [EPR25340-17] (ab289963)
      All lanes : Anti-E2F6 antibody [EPR25340-17] (ab289963) at 1/1000 dilution

      Lane 1 : Mouse E17 brain tissue lysate
      Lane 2 : Jurkat (human T cell leukemia T lymphocyte) whole cell lysate
      Lane 3 : K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate
      Lane 4 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
      Lane 5 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

      Predicted band size: 32 kDa
      Observed band size: 35 kDa why is the actual band size different from the predicted?



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

      The expression profile/molecular weight observed is consistent with that described in the literature (PMID: 28964969).

      There is an unknown band at ~100kDa.

      Exposure time: 10 seconds

       

    • Flow Cytometry (Intracellular) - Anti-E2F6 antibody [EPR25340-17] (ab289963)
      Flow Cytometry (Intracellular) - Anti-E2F6 antibody [EPR25340-17] (ab289963)

      Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized Jurkat (Human T cell leukemia T lymphocyte) cells labelling E2F6 with ab289963 at 1/500 dilution (0.1µg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

    • Immunocytochemistry/ Immunofluorescence - Anti-E2F6 antibody [EPR25340-17] (ab289963)
      Immunocytochemistry/ Immunofluorescence - Anti-E2F6 antibody [EPR25340-17] (ab289963)

      Immunofluorescent analysis of 80% Methanol-fixed, 0.1% TritonX-100 permeabilized Jurkat cells labeling E2F6 with ab289963 at 1/100 dilution, followed by ab150081 Goat Anti-Rbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in Jurkat cell line. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

      Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E2F6 antibody [EPR25340-17] (ab289963)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E2F6 antibody [EPR25340-17] (ab289963)

      Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labelling E2F6 with ab289963 at 1/5000 dilution, followed by LeicaDS9800 (Bond®, Polymer Refine Detection). Nuclear staining on mouse cerebrum. The section was incubated with ab289963 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is LeicaDS9800 (Bond®, Polymer Refine Detection).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 10 mins.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E2F6 antibody [EPR25340-17] (ab289963)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-E2F6 antibody [EPR25340-17] (ab289963)

      Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labelling E2F6 with ab289963 at 1/5000 dilution, followed by LeicaDS9800 (Bond , Polymer Refine Detection). Nuclear staining on human cerebrum. The section was incubated with ab289963 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

      Secondary antibody only control: Secondary antibody is LeicaDS9800 (Bond®, Polymer Refine Detection).

      Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 10 mins.

    • Immunoprecipitation - Anti-E2F6 antibody [EPR25340-17] (ab289963)
      Immunoprecipitation - Anti-E2F6 antibody [EPR25340-17] (ab289963)

      E2F6 was immunoprecipitated from NIH/3T3 (mouse embryonic fibroblast) whole cell lysate with ab289963 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab289963 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

      Lane 1: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate10 µg

      Lane 2: ab289963 IP in NIH/3T3 whole cell lysate

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab289963 in NIH/3T3 whole cell lysate

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 3.25 seconds

       

    • Western blot - Anti-E2F6 antibody [EPR25340-17] (ab289963)
      Western blot - Anti-E2F6 antibody [EPR25340-17] (ab289963)
      All lanes : Anti-E2F6 antibody [EPR25340-17] (ab289963) at 1/1000 dilution

      Lane 1 : MEF (mouse embryonic fibroblast (immortalized)) whole cell lysate
      Lane 2 : NIH/3T3 (mouse embryonic fibroblast) whole cell lysate
      Lane 3 : C2C12 (mouse myoblasts myoblast) whole cell lysate
      Lane 4 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
      Lane 5 : PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

      Predicted band size: 32 kDa
      Observed band size: 35 kDa why is the actual band size different from the predicted?



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

      The expression profile/molecular weight observed is consistent with that described in the literature (PMID: 15936277).

      Exposure time: 10 seconds

       

    • Anti-E2F6 antibody [EPR25340-17] (ab289963)
      Anti-E2F6 antibody [EPR25340-17] (ab289963)

    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 (0)

    ab289963 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。

    ab289963 は論文での使用が確認できていません。

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