Overview

  • Product name
    Anti-CD40 antibody
    See all CD40 primary antibodies
  • Description
    Rabbit polyclonal to CD40
  • Host species
    Rabbit
  • Specificity
    The antibody specificity is confirmed by specific peptide inhibition in immunoblotting studies.
  • Tested applications
    Suitable for: IHC-P, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Sheep, Rabbit, Guinea pig, Hamster, Cow, Human, Pig, Monkey
    Predicted to work with: Chicken, Dog
  • Immunogen

    Synthetic peptide:

    CQPVTQEDGKESRISVQERQ

    conjugated to KLH, corresponding to amino acids 258-277 of Human CD40.

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • Positive control
    • HeLa Cell Lysate. This antibody gave a positive result in IHC in the following FFPE tissue: Human normal tonsil.
  • General notes
    Storage below -20°C is not recommended. For maximum product recovery centrifuge the product vial before removing cap. Shipped on gel packs.

Properties

Applications

Our Abpromise guarantee covers the use of ab13545 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
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 40 kDa (predicted molecular weight: 34 kDa).

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.
  • Information by UniProt
  • Database links
    • 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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody (ab13545)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody (ab13545)

      IHC image of CD40 staining in Human normal tonsil formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ 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 ab13545, 1µg/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.

    • Western blot - Anti-CD40 antibody (ab13545)
      Western blot - Anti-CD40 antibody (ab13545)
      Anti-CD40 antibody (ab13545) at 1/1000 dilution + Recombinant human CD40 protein (Fc Chimera Active) (ab83917) at 1 µg

      Secondary
      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: 34 kDa


      Exposure time: 8 minutes
    • Western blot - Anti-CD40 antibody (ab13545)
      Western blot - Anti-CD40 antibody (ab13545)
      All lanes : Anti-CD40 antibody (ab13545) at 1/200 dilution

      Lane 1 : ESK4 cell lysate
      Lane 2 : Molt 4 cell lysate

      Predicted band size: 34 kDa

    • Western blot - Anti-CD40 antibody (ab13545)
      Western blot - Anti-CD40 antibody (ab13545)
      All lanes : Anti-CD40 antibody (ab13545) at 1/1000 dilution

      Lanes 1-2 : Control human HCAEC cells - whole cell lysate
      Lanes 3-4 : Treated human HCAEC cells - whole cell lysate

      Secondary
      All lanes : HRP conjugated Goat Anti-Rabbit IgG

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 34 kDa
      Observed band size: 45 kDa (why is the actual band size different from the predicted?)


      Exposure time: 20 seconds


      This image is courtesy of an Abreview submitted on 10 August 2005.

      See Abreview

    • Western blot - Anti-CD40 antibody (ab13545)
      Western blot - Anti-CD40 antibody (ab13545)This image is courtesy of an anonymous Abreview
      All lanes : Anti-CD40 antibody (ab13545) at 1/5000 dilution

      Lane 1 : Human B chronic lymphocytic leukaemia cells, nuclear fraction
      Lane 2 : Human B chronic lymphocytic leukaemia cells, cytoskeletal fraction
      Lane 3 : Human B chronic lymphocytic leukaemia cells, membrane fraction
      Lane 4 : Human B chronic lymphocytic leukaemia cells, cytosolic fraction

      Lysates/proteins at 18 µg per lane.

      Secondary
      All lanes : HRP conjugated goat anti-rabbit

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 34 kDa
      Observed band size: 42 kDa (why is the actual band size different from the predicted?)


      Exposure time: 3 minutes


      The primary antibody was incubated with the membrane for 16 hours at 4°C.

      See Abreview

    References

    This product has been referenced in:
    • Shoji T  et al. Local convection-enhanced delivery of an anti-CD40 agonistic monoclonal antibody induces antitumor effects in mouse glioma models. Neuro Oncol 18:1120-8 (2016). Read more (PubMed: 26917236) »
    • Qiu X  et al. CD40 ligand induces RIP1-dependent, necroptosis-like cell death in low-grade serous but not serous borderline ovarian tumor cells. Cell Death Dis 6:e1864 (2015). IHC-P ; Human . Read more (PubMed: 26313915) »

    See all 5 Publications for this product

    Publishing research using ab13545? Please let us know so that we can cite the reference in this datasheet.

    Customer reviews and Q&As

    Submit a review Submit a question

    7 reviews or Q&A

    10
    Votes: Highest First

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD40 antibody

     Inconclusive
    Abreviews
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Brain tissue)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate (0.01M) pH=6
    Permeabilization
    Yes - 0.05% Tween
    Specification
    Brain tissue
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 1% · Temperature: 21°C
    Fixative
    Paraformaldehyde
    Read More
    abreview image
    Username

    Boyd Kenkhuis

    Verified customer

    Submitted Aug 22 2018

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD40 antibody

     Good
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (spleen)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9.0
    Permeabilization
    No
    Specification
    spleen
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Fixative
    10% NBF
    Read More
    abreview image
    Username

    Abcam user community

    Verified customer

    Submitted Dec 28 2017

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD40 antibody

     Good
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (skin)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9.0
    Permeabilization
    No
    Specification
    skin
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Fixative
    Formaldehyde
    Read More
    abreview image
    Username

    Abcam user community

    Verified customer

    Submitted Dec 27 2017

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD40 antibody

     Excellent
    Abreviews
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (human tonsil)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Dako Low pH
    Permeabilization
    No
    Specification
    human tonsil
    Blocking step
    Flex endogenouse peroxidase block as blocking agent for 5 minute(s) · Concentration: 100% · Temperature: RT°C
    Fixative
    Paraformaldehyde
    Read More
    abreview image
    Username

    Abcam user community

    Verified customer

    Submitted Nov 19 2015

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD40 antibody

     Excellent
    Abreviews
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Blocking step
    Dako Flex peroxidase as blocking agent for 5 minute(s) · Concentration: 100% · Temperature: RT°C
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Low pH
    Sample
    Human Tissue sections (Malignant melanoma/lymphocytes)
    Specification
    Malignant melanoma/lymphocytes
    Permeabilization
    No
    Fixative
    Formaldehyde
    Read More
    Username

    Abcam user community

    Verified customer

    Submitted Jan 20 2015

    Western blot abreview for Anti-CD40 antibody

     Good
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - other (Human B chronic lymphocytic leukaemia cells)
    Loading amount
    18 µg
    Specification
    Human B chronic lymphocytic leukaemia cells
    Gel Running Conditions
    Reduced Denaturing (12% Tris-Glycine Gel)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
    Read More
    abreview image
    Username

    Abcam user community

    Verified customer

    Submitted Feb 22 2008

    Western blot abreview for Anti-CD40 antibody

     Excellent
    Abreviews
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (HCAEC)
    Specification
    HCAEC
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
    Read More
    abreview image
    Username

    Abcam user community

    Verified customer

    Submitted Aug 10 2005

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