Overview

  • Product name
  • Description
    Rabbit polyclonal to CD40
  • 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.

  • 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

  • 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.

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

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

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

    See 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
    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

    This image is courtesy of an anonymous Abreview

    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

Customer reviews and Q&As

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
Username

Abcam user community

Verified customer

Submitted Nov 19 2015

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
Username

Abcam user community

Verified customer

Submitted Jan 20 2015

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
Username

Abcam user community

Verified customer

Submitted Feb 22 2008

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%
Username

Abcam user community

Verified customer

Submitted Aug 10 2005

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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