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  1. Link

    cd40-antibody-ab113701.pdf

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Immunology Adaptive Immunity B Cells CD
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Validated using a knockout cell line

Anti-CD40 antibody (ab113701)

  • Datasheet
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Western blot - Anti-CD40 antibody (ab113701)
  • Western blot - Anti-CD40 antibody (ab113701)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody (ab113701)

Key features and details

  • Rabbit polyclonal to CD40
  • Suitable for: WB, IHC-P
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG

Get better batch-to-batch reproducibility with a recombinant antibody

Product image
Anti-CD40 antibody [EPR20540] (ab213205)
  • Research with confidence – consistent and reproducible results with every batch
  • Long-term and scalable supply – powered by recombinant technology for fast production
  • Success from the first experiment – confirmed specificity through extensive validation
  • Ethical standards compliant – production is animal-free

Overview

  • Product name

    Anti-CD40 antibody
    See all CD40 primary antibodies
  • Description

    Rabbit polyclonal to CD40
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Macaque monkey, Gorilla
  • Immunogen

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

  • Positive control

    • WB: U-2 OS and Raji cell lysates; Human spleen and thymus tissue lysates. IHC-FFPE: Human spleen tissue.
  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Adaptive Immunity
    • B Cells
    • CD
    • Cardiovascular
    • Atherosclerosis
    • Vascular Inflammation
    • Innate and adaptive immunity
    • Stem Cells
    • Hematopoietic Progenitors
    • Lymphoid
    • B Lymphocytic Lineage
    • Stem Cells
    • Hematopoietic Progenitors
    • Myeloid
    • Dendritic Cell Lineage
    • Neuroscience
    • Processes

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • KO cell lines

    • Human CD40 knockout U-2 OS cell line (ab262486)
  • KO cell lysates

    • Human CD40 knockout U-2 OS cell lysate (ab263923)
  • Recombinant Protein

    • Recombinant human CD40 protein (Active) (ab155710)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab113701 in the following tested applications.

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

Application Abreviews Notes
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 43 kDa (predicted molecular weight: 31 kDa).
IHC-P
Use a concentration of 5 µg/ml.
Notes
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 43 kDa (predicted molecular weight: 31 kDa).
IHC-P
Use a concentration of 5 µg/ml.

Target

  • Function

    Receptor for TNFSF5/CD40LG.
  • Tissue specificity

    B-cells and in primary carcinomas.
  • Involvement in disease

    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.
  • Sequence similarities

    Contains 4 TNFR-Cys repeats.
  • Cellular localization

    Secreted and Cell membrane.
  • Target information above from: UniProt accession P25942 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 958 Human
    • Omim: 109535 Human
    • SwissProt: P25942 Human
    • Unigene: 472860 Human
    • Alternative names

      • 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

    Images

    • Western blot - Anti-CD40 antibody (ab113701)
      Western blot - Anti-CD40 antibody (ab113701)
      All lanes : Anti-CD40 antibody (ab113701) at 1 µg/ml

      Lane 1 : Wild-type U-2 OS cell lysate
      Lane 2 : CD40 knockout U-2 OS cell lysate
      Lane 3 : Raji cell lysate

      Lysates/proteins at 40 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 31 kDa
      Observed band size: 45 kDa why is the actual band size different from the predicted?



      Lanes 1 - 3: Merged signal (red and green). Green - ab113701 observed at 45 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.

      ab113701 was shown to react with CD40 in wild-type U-2 OS cells in Western blot. Loss of signal was observed when CD40 knockout cell line ab262486 (knockout cell lysate ab263923) was used. Wild-type and CD40 knockout U-2 OS cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab113701 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at 1 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 hour at room temperature before imaging.

    • Western blot - Anti-CD40 antibody (ab113701)
      Western blot - Anti-CD40 antibody (ab113701)
      All lanes : Anti-CD40 antibody (ab113701) at 1 µg/ml

      Lane 1 : Human spleen tissue lysate - total protein (ab29699)
      Lane 2 : Human thymus tissue lysate - total protein (ab30146)

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 31 kDa
      Observed band size: 43 kDa why is the actual band size different from the predicted?
      Additional bands at: 105 kDa, 41 kDa (possible cleavage fragment). We are unsure as to the identity of these extra bands.


      Exposure time: 10 minutes


      CD40 contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted. The predicted molecular weight of CD40 is 31 kDa (SwissProt), however we expect to observe a banding pattern around 43 kDa. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody (ab113701)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody (ab113701)

      IHC image of CD40 staining in Human normal spleen formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab113701, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    Protocols

    • Western blot protocols
    • Immunohistochemistry protocols

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (0)

    Publishing research using ab113701? Please let us know so that we can cite the reference in this datasheet.

    ab113701 has not yet been referenced specifically in any publications.

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