Recombinant Anti-CD40 antibody [EPR20540] (ab213205)
![Western blot - Anti-CD40 antibody [EPR20540] (ab213205)](https://www.abcam.com/ps/products/213/ab213205/Images/ab213205-380877-anti-cd40-antibody-epr20540-western-blot-u2-os-wt-u2-os-cd40-knockout-raji-mcf7.jpg "Western blot - Anti-CD40 antibody [EPR20540] (ab213205)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20540] to CD40
- Suitable for: IHC-P, WB
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-CD40 antibody [EPR20540]
See all CD40 primary antibodies -
Description
Rabbit monoclonal [EPR20540] to CD40 -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, WBmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Human tonsil, lymph node and lymphoma lysates; Raji, U-2 OS, and Daudi whole cell lysates. IHC-P: Human tonsil and large B cell lymphoma tissues.
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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: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20540 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab213205 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 |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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| WB |
1/2000. Detects a band of approximately 42 kDa (predicted molecular weight: 30 kDa).
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| Notes |
|---|
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IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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WB
1/2000. Detects a band of approximately 42 kDa (predicted molecular weight: 30 kDa). |
Target
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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
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Database links
- Entrez Gene: 958 Human
- Omim: 109535 Human
- SwissProt: P25942 Human
- Unigene: 472860 Human
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Alternative names
- AI326936 antibody
- B cell associated molecule CD40 antibody
- B cell surface antigen CD40 antibody
see all
Images
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Western blot - Anti-CD40 antibody [EPR20540] (ab213205)All lanes : Anti-CD40 antibody [EPR20540] (ab213205) at 1/2000 dilution
Lane 1 : Wild-type U-2 OS whole cell lysate
Lane 2 : CD40 knockout U-2 OS whole cell lysate
Lane 3 : Raji (Human Burkitt's lymphoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 30 kDa
Observed band size: 42 kDa why is the actual band size different from the predicted?Lanes 1 - 3: Merged signal (red and green). Green - ab213205 observed at 42 kDa. Red - loading control, ab7291 (Mouse anti-Alpha Tubulin [DM1A] observed at 55kDa.
ab213205 was shown to react with CD40 in U-2 OS wild-type cells in Western blot. Loss of signal was observed when CD40 knockout sample was used. U-2 OS wild-type and CD40 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk in TBS-T (0.1% Tween®) before incubation with ab213205 and ab7291 (Mouse anti-Alpha Tubulin [DM1A] overnight at 4°C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were developed 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.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR20540] (ab213205)](https://www.abcam.com/ps/products/213/ab213205/Images/ab213205-274824-anti-cd40-antibody-epr20540-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR20540] (ab213205)Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD40 with ab213205 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Membranous and cytoplasmic staining on germinal center of human tonsil is observed [PMID: 10360965] [PMID: 7507299] [PMID:24452203].
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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![Western blot - Anti-CD40 antibody [EPR20540] (ab213205)](https://www.abcam.com/ps/products/213/ab213205/Images/ab213205-274825-anti-cd40-antibody-epr20540-western-blot.jpg)
Western blot - Anti-CD40 antibody [EPR20540] (ab213205)All lanes : Anti-CD40 antibody [EPR20540] (ab213205) at 1/2000 dilution
Lane 1 : Human tonsil lysate
Lane 2 : Human lymph node lysate
Lane 3 : Human lymphoma lysate
Lane 4 : Raji (Human Burkitt's lymphoma cell line) whole cell lysate
Lane 5 : Daudi (Human Burkitt's lymphoma cell line) whole cell lysate
Lane 6 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
Lanes 1-3 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution
Lanes 4-6 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 30 kDa
Observed band size: 42 kDa why is the actual band size different from the predicted?Blocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1,2,3 and 4: 30 seconds; Lane 5,6 and 7: 5 seconds.
The molecular weight observed is consistent with the literature (PMID: 23404288);
Negative control: Jurkat (PMID:10498643; PMID:15708600).
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR20540] (ab213205)](https://www.abcam.com/ps/products/213/ab213205/Images/ab213205-274823-anti-cd40-antibody-epr20540-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR20540] (ab213205)Immunohistochemical analysis of paraffin-embedded human large B cell lymphoma tissue labeling CD40 with ab213205 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Membranous staining on human large B cell lymphoma is observed [PMID: 7507299].
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR20540] (ab213205)](https://www.abcam.com/ps/products/213/ab213205/Images/ab213205-6-anti-cd40-antibody-epr20540-tissue-microarray-human-ab213205.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR20540] (ab213205)Tissue Microarrays stained for "Anti-CD40 antibody [EPR20540]” using "ab213205" in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). The sections were incubated with ab213205 at +4°C overnight followed by Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/500.
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![Anti-CD40 antibody [EPR20540] (ab213205)](https://www.abcam.com/ps/products/213/ab213205/Images/ab213205-5-benefits-of-recombinant-antibodies.png)
Anti-CD40 antibody [EPR20540] (ab213205)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (0)
ab213205 has not yet been referenced specifically in any publications.