遺伝子ノックアウト細胞株 検証済リコンビナントRabMAb

Anti-CD40 抗体 [EPR20735] (ab224639)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR20735] (ab224639)
  • Immunocytochemistry/ Immunofluorescence - Anti-CD40 antibody [EPR20735] (ab224639)
  • Flow Cytometry - Anti-CD40 antibody [EPR20735] (ab224639)
  • Immunocytochemistry/ Immunofluorescence - Anti-CD40 antibody [EPR20735] (ab224639)
  • Western blot - Anti-CD40 antibody [EPR20735] (ab224639)
  • Immunoprecipitation - Anti-CD40 antibody [EPR20735] (ab224639)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR20735] (ab224639)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR20735] (ab224639)
  • Anti-CD40 antibody [EPR20735] (ab224639)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20735] to CD40
  • Suitable for: WB, IHC-P, Flow Cyt, IP, ICC/IF
  • Knockout validated
  • Reacts with: Human

製品の概要

  • 製品名

    Anti-CD40 antibody [EPR20735]
    CD40 一次抗体 製品一覧

  • 製品の詳細

    Rabbit monoclonal [EPR20735] to CD40

  • 由来種

    Rabbit

  • アプリケーション

    適用あり: WB, IHC-P, Flow Cyt, IP, 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: Human lymph node and lymphoma tissue lysates; Raji whole cell lysate. IHC-P: Human tonsil, Hodgkin's lymphoma and diffuse large B-cell lymphoma tissues. ICC/IF: Raji cells. Flow Cyt: Raji cells. IP: Raji whole cell lysate. ICC/IF KO: U2OS cells (U2OS-CD40 knockout cells used as negative cell line)

  • 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ab224639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 42 kDa (predicted molecular weight: 30 kDa).
IHC-P
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt
1/600.

ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IP
1/30.
ICC/IF
1/100.
特記事項
WB
1/1000. Detects a band of approximately 42 kDa (predicted molecular weight: 30 kDa).
IHC-P
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt
1/600.

ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IP
1/30.
ICC/IF
1/100.

ターゲット情報

  • 機能

    Receptor for TNFSF5/CD40LG.

  • 組織特異性

    B-cells and in primary carcinomas.

  • 関連疾患

    Defects in CD40 are the cause of hyper-IgM immunodeficiency syndrome type 3 (HIGM3) [MIM:606843]; also known as hyper-IgM syndrome 3. HIGM3 is an autosomal recessive disorder which includes an inability of B cells to undergo isotype switching, one of the final differentiation steps in the humoral immune system, an inability to mount an antibody-specific immune response, and a lack of germinal center formation.

  • 配列類似性

    Contains 4 TNFR-Cys repeats.

  • 細胞内局在

    Secreted and Cell membrane.
  • Information by UniProt

  • 参照データベース

    • 別名

      • AI326936 antibody
      • B cell associated molecule CD40 antibody
      • B cell surface antigen CD40 antibody
      • B cell-associated molecule antibody
      • B-cell surface antigen CD40 antibody
      • Bp50 antibody
      • CD 40 antibody
      • CD40 antibody
      • CD40 antigen (TNF receptor superfamily member 5) antibody
      • CD40 antigen antibody
      • CD40 molecule antibody
      • CD40 molecule, TNF receptor superfamily member 5 antibody
      • CD40 protein antibody
      • CD40 type II isoform antibody
      • CD40L receptor antibody
      • CDw40 antibody
      • GP39 antibody
      • HIGM1 antibody
      • IGM antibody
      • IMD3 antibody
      • MGC9013 antibody
      • Nerve growth factor receptor related B lymphocyte activation molecule antibody
      • OTTHUMP00000031699 antibody
      • OTTHUMP00000031700 antibody
      • p50 antibody
      • T-BAM antibody
      • TBAM antibody
      • TNF receptor superfamily member 5 antibody
      • TNFRSF5 antibody
      • TNR5_HUMAN antibody
      • TRAP antibody
      • Tumor necrosis factor receptor superfamily , member 5 antibody
      • Tumor necrosis factor receptor superfamily member 5 antibody
      • Tumor necrosis factor receptor superfamily member 5 precursor antibody
      • Tumor necrosis factor receptor superfamily, member 5, isoform CRA_a antibody
      see all

    画像

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR20735] (ab224639)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR20735] (ab224639)

      Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD40 with ab224639 at 1/250 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous and cytoplasmic staining in human tonsil tissue (PMID:20593977; PMID:7507299) is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

      Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Immunocytochemistry/ Immunofluorescence - Anti-CD40 antibody [EPR20735] (ab224639)
      Immunocytochemistry/ Immunofluorescence - Anti-CD40 antibody [EPR20735] (ab224639)

      ab224639 staining CD40 in wild-type U2OS cells (top panel) and CD40 knockout U2OS cells (bottom panel). The cells were fixed with PFA (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab224639 at 1/100 dilution and ab7291 (Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in pseudo color red).  Nuclear DNA was labelled in blue with DAPI.

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    • Flow Cytometry - Anti-CD40 antibody [EPR20735] (ab224639)
      Flow Cytometry - Anti-CD40 antibody [EPR20735] (ab224639)

      Flow cytometric analysis of Raji (human Burkitt's lymphoma cell line) cell line labeling CD40 with ab224639 at 1/600 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody.

      Gated on total viable cells.

    • Immunocytochemistry/ Immunofluorescence - Anti-CD40 antibody [EPR20735] (ab224639)
      Immunocytochemistry/ Immunofluorescence - Anti-CD40 antibody [EPR20735] (ab224639)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Raji (human Burkitt's lymphoma cell line) cells labeling CD40 with ab224639 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous and weakly cytoplasmic staining in Raji cell line.

      Negative control: Jurkat T cell line (PMID:10498643).

      The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).

      Secondary antibody only control: PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    • Western blot - Anti-CD40 antibody [EPR20735] (ab224639)
      Western blot - Anti-CD40 antibody [EPR20735] (ab224639)
      All lanes : Anti-CD40 antibody [EPR20735] (ab224639) at 1/1000 dilution

      Lane 1 : Human lymph node tissue lysate
      Lane 2 : Human lymphoma tissue lysate
      Lane 3 : Raji (human Burkitt's lymphoma cell line) whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Developed using the ECL technique.

      Predicted band size: 30 kDa
      Observed band size: 42 kDa why is the actual band size different from the predicted?


      Exposure time: 15 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Immunoprecipitation - Anti-CD40 antibody [EPR20735] (ab224639)
      Immunoprecipitation - Anti-CD40 antibody [EPR20735] (ab224639)

      CD40 was immunoprecipitated from 0.35 mg of Raji (human Burkitt's lymphoma cell line) whole cell lysate with ab224639 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab224639 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

      Lane 1: Raji whole cell lysate 10 μg (Input).
      Lane 2: ab224639 IP in Raji whole cell lysate (+).
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab224639 in Raji whole cell lysate (-).

      Exposure time: 3 minutes.

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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR20735] (ab224639)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR20735] (ab224639)

      Immunohistochemical analysis of paraffin-embedded human diffuse large B-cell lymphoma tissue labeling CD40 with ab224639 at 1/250 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous staining in human diffuse large B-cell lymphoma tissue (PMID: 20593977) is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

      Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR20735] (ab224639)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR20735] (ab224639)

      Immunohistochemical analysis of paraffin-embedded human Hodgkin's lymphoma tissue labeling CD40 with ab224639 at 1/250 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous and cytoplasmic staining in Reed-Sternberg cells and tumor cells of human Hodgkin's lymphoma (PMID:7507299) is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

      Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Anti-CD40 antibody [EPR20735] (ab224639)
      Anti-CD40 antibody [EPR20735] (ab224639)

    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 (6)

    ab224639 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。

    ab224639 は 6 報の論文で使用されています。

    • Luo C  et al. CD40 is Positively Correlated with the Expression of Nucleophosmin in Cisplatin-Resistant Bladder Cancer. J Oncol 2020:3676751 (2020). PubMed: 32411234
    • Zhou ZQ  et al. Adipose extracellular matrix promotes skin wound healing by inducing the differentiation of adipose-derived stem cells into fibroblasts. Int J Mol Med 43:890-900 (2019). PubMed: 30535488
    • Wang L  et al. Identification of circular RNA Hsa_circ_0001879 and Hsa_circ_0004104 as novel biomarkers for coronary artery disease. Atherosclerosis 286:88-96 (2019). PubMed: 31103880
    • Jiang C  et al. CRISPR/Cas9 Screens Reveal Multiple Layers of B cell CD40 Regulation. Cell Rep 28:1307-1322.e8 (2019). PubMed: 31365872
    • Hong D  et al. Human-induced pluripotent stem cell-derived macrophages and their immunological function in response to tuberculosis infection. Stem Cell Res Ther 9:49 (2018). WB . PubMed: 29482598
    • Journo G  et al. Modulation of Cellular CpG DNA Methylation by Kaposi's Sarcoma-Associated Herpesvirus. J Virol 92:N/A (2018). PubMed: 29899086

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