Recombinant Anti-CD40 Ligand antibody [EPR26530-17] (ab303610)
![Western blot - Anti-CD40 Ligand antibody [EPR26530-17] (ab303610)](https://www.abcam.com/ps/products/303/ab303610/Images/ab303610-498334-anti-cd40-ligand-antibody-epr26530-17-western-blot-untreated-jurkat-human-t-cell-leukemia-t-lymphocyte-from-peripheral-blood-whole-cell-lysate-50-ug.jpg "Western blot - Anti-CD40 Ligand antibody [EPR26530-17] (ab303610)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR26530-17] to CD40L
- Suitable for: WB, IHC-P, IP
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-CD40 Ligand antibody [EPR26530-17]
See all CD40L primary antibodies -
Description
Rabbit monoclonal [EPR26530-17] to CD40L -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, IPmore details
Unsuitable for: Flow Cyt or ICC/IF -
Species reactivity
Reacts with: Human
Does not react with: Mouse, Rat -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: PMA/BFA/monensin treated Jurkat whole cell lysate. D1.1 whole cell lysate. Human lymphoma, spleen and tonsil tissue lysate. IHC-P: Human tonsil tissue. PMA/BFA/monensin treated Jurkat cell pellet. IP: D1.1 whole cell lysate.
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General notes
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR26530-17 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab303610 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 |
1/1000. Predicted molecular weight: 29 kDa.
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| IHC-P |
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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| IP |
1/30.
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| Notes |
|---|
|
WB
1/1000. Predicted molecular weight: 29 kDa. |
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IHC-P
1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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IP
1/30. |
Target
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Function
Mediates B-cell proliferation in the absence of co-stimulus as well as IgE production in the presence of IL-4. Involved in immunoglobulin class switching.
Release of soluble CD40L from platelets is partially regulated by GP IIb/IIIa, actin polymerization, and an matrix metalloproteinases (MMP) inhibitor-sensitive pathway. -
Tissue specificity
Specifically expressed on activated CD4+ T-lymphocytes. -
Involvement in disease
Defects in CD40LG are the cause of X-linked immunodeficiency with hyper-IgM type 1 (HIGM1) [MIM:308230]; also known as X-linked hyper IgM syndrome (XHIM). HIGM1 is an immunoglobulin isotype switch defect characterized by elevated concentrations of serum IgM and decreased amounts of all other isotypes. Affected males present at an early age (usually within the first year of life) recurrent bacterial and opportunistic infections, including Pneumocystis carinii pneumonia and intractable diarrhea due to cryptosporidium infection. Despite substitution treatment with intravenous immunoglobulin, the overall prognosis is rather poor, with a death rate of about 10% before adolescence. -
Sequence similarities
Belongs to the tumor necrosis factor family. -
Post-translational
modificationsThe soluble form derives from the membrane form by proteolytic processing.
N-linked glycan is a mixture of high mannose and complex type. Glycan structure does not influence binding affinity to CD40.
Not O-glycosylated. -
Cellular localization
Secreted and Cell membrane. - Information by UniProt
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Database links
- Entrez Gene: 959 Human
- Omim: 300386 Human
- SwissProt: P29965 Human
- Unigene: 592244 Human
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Alternative names
- CD154 antibody
- CD40 antigen ligand antibody
- CD40 ligand antibody
see all
Images
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Western blot - Anti-CD40 Ligand antibody [EPR26530-17] (ab303610)All lanes : Anti-CD40 Ligand antibody [EPR26530-17] (ab303610) at 1/1000 dilution
Lane 1 : Untreated Jurkat (human T cell leukemia T lymphocyte from peripheral blood), whole cell lysate
Lane 2 : Jurkat treated with /ml 48h PMA for 48h, then treated with 300ng/ml BFA and 3uM monensin for 24h, whole cell lysate
Lane 3 : D1.1 (human T lymphoblast), whole cell lysate
Lane 4 : Raji (human Burkitt's lymphoma B lymphocyte), whole cell lysate
Lysates/proteins at 50 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/1000 dilution
Predicted band size: 29 kDa
Observed band size: 17,25-30 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
Negative control: Raji (PMID:12805559)
The antibody recognizes both membrane bound (25-30KDa) and soluble (17KDa) form.
Exposure time: 3.25 seconds
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![Western blot - Anti-CD40 Ligand antibody [EPR26530-17] (ab303610)](https://www.abcam.com/ps/products/303/ab303610/Images/ab303610-498336-anti-cd40-ligand-antibody-epr26530-17-western-blot-human-lymphma-tissue-lysate-50-ug.jpg)
Western blot - Anti-CD40 Ligand antibody [EPR26530-17] (ab303610)All lanes : Anti-CD40 Ligand antibody [EPR26530-17] (ab303610) at 1/1000 dilution
Lane 1 : Human lymphoma tissue lysate
Lane 2 : Human spleen tissue lysate
Lane 3 : Human tonsil tissue lysate
Lane 4 : Human skeletal muscle tissue lysate
Lysates/proteins at 50 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/1000 dilution
Predicted band size: 29 kDa
Observed band size: 17,25-30 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
Negative control: human skeletal muscle (PMID:1281209)
The antibody recognizes both membrane bound (25-30KDa) and soluble (17KDa) form.
Exposure time: 180 seconds
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 Ligand antibody [EPR26530-17] (ab303610)](https://www.abcam.com/ps/products/303/ab303610/Images/ab303610-2-anti-cd40-ligand-antibody-epr26530-17-immunohistochemistry-human-tonsil.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 Ligand antibody [EPR26530-17] (ab303610)Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD40 Ligand with ab303610 at 1/500 dilution (1.062 µg/ml) followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on human tonsil (PMID: 7699321).
The section was incubated with ab303610 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 ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 Ligand antibody [EPR26530-17] (ab303610)](https://www.abcam.com/ps/products/303/ab303610/Images/ab303610-3-anti-cd40-ligand-antibody-epr26530-17-immunohistochemistry-a-wild-type-jurkat.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 Ligand antibody [EPR26530-17] (ab303610)Immunohistochemical analysis of paraffin-embedded cell pellets labeling CD40 Ligand with ab303610 at 1/500 dilution (1.062 µg/ml) followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Positive staining on wild-type Jurkat (human T cell leukemia T lymphocyte from peripheral blood) cell pellet (A) and Jurkat treated with 50ng/ml 48h PMA for 48h, then treated with 300ng/ml BFA and 3µM monensin for 24h cell pellet (B). No staining on wild-type Raji (human Burkitt's lymphoma B lymphocyte) cell pellet (C).
The section was incubated with ab303610 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 Ligand antibody [EPR26530-17] (ab303610)](https://www.abcam.com/ps/products/303/ab303610/Images/ab303610-4-anti-cd40-ligand-antibody-epr26530-17-immunohistochemistry-human-skeletal-muscl.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 Ligand antibody [EPR26530-17] (ab303610)Immunohistochemical analysis of paraffin-embedded human skeletal muscle tissue labeling CD40 Ligand with ab303610 at 1/500 dilution (1.062 µg/ml) followed by ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Negative control: No staining on human skeletal muscle (PMID: 1281209).
The section was incubated with ab303610 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 ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).
Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.
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![Immunoprecipitation - Anti-CD40 Ligand antibody [EPR26530-17] (ab303610)](https://www.abcam.com/ps/products/303/ab303610/Images/ab303610-1-anti-cd40-ligand-antibody-epr26530-17-immunoprecipitation-d11.jpg)
Immunoprecipitation - Anti-CD40 Ligand antibody [EPR26530-17] (ab303610)CD40 Ligand was immunoprecipitated from 0.35 mg D1.1 (human T lymphoblast) whole cell lysate 10 µg with ab303610 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab303610 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: D1.1 whole cell lysate 10 µg
Lane 2: ab303610 IP in D1.1 whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab303610 in D1.1 whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST
Exposure time: 180 seconds
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![Anti-CD40 Ligand antibody [EPR26530-17] (ab303610)](https://www.abcam.com/ps/products/303/ab303610/Images/ab303610-498273-benefitsofrecombinantantibodies.jpg)
Anti-CD40 Ligand antibody [EPR26530-17] (ab303610)
Protocols
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.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab303610 has not yet been referenced specifically in any publications.