Recombinant Anti-CD40 antibody [EPR20735] (ab224639)
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR20735] (ab224639)](https://www.abcam.com/ps/products/224/ab224639/Images/ab224639-293852-anti-cd40-antibody-epr20735-immunohistochemistry.jpg "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR20735] (ab224639)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20735] to CD40
- Suitable for: WB, IHC-P, Flow Cyt, IP, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
-
Product name
Anti-CD40 antibody [EPR20735]
See all CD40 primary antibodies -
Description
Rabbit monoclonal [EPR20735] to CD40 -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, Flow Cyt, IP, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
-
Positive control
- WB: Human lymph node and lymphoma tissue lysates; Raji whole cell lysate. IHC-P: Human tonsil, Hodgkin's lymphoma and diffuse large B-cell lymphoma tissues. ICC/IF: Raji cells. Flow Cyt: Raji cells. IP: Raji whole cell lysate. ICC/IF KO: U2OS cells (U2OS-CD40 knockout cells used as negative cell line)
-
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
-
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: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20735 -
Isotype
IgG -
Research areas
Associated products
-
Alternative Versions
-
Compatible Secondaries
-
Conjugation kits
-
Isotype control
-
KO cell lines
-
Recombinant Protein
-
Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab224639 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. Detects a band of approximately 42 kDa (predicted molecular weight: 30 kDa).
|
|
| IHC-P |
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
|
| Flow Cyt |
1/600.
ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
|
| IP |
1/30.
|
|
| ICC/IF |
1/100.
|
| Notes |
|---|
|
WB
1/1000. Detects a band of approximately 42 kDa (predicted molecular weight: 30 kDa). |
|
IHC-P
1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
|
Flow Cyt
1/600. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
|
IP
1/30. |
|
ICC/IF
1/100. |
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
- 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
see all
Images
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR20735] (ab224639)
Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling CD40 with ab224639 at 1/250 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous and cytoplasmic staining in human tonsil tissue (PMID:20593977; PMID:7507299) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
-
![Immunocytochemistry/ Immunofluorescence - Anti-CD40 antibody [EPR20735] (ab224639)](https://www.abcam.com/ps/products/224/ab224639/Images/ab224639-381128-ICCIFab224639GR3222715201.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-CD40 antibody [EPR20735] (ab224639)ab224639 staining CD40 in wild-type U2OS cells (top panel) and CD40 knockout U2OS cells (bottom panel). The cells were fixed with PFA (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab224639 at 1/100 dilution and ab7291 (Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
-
![Flow Cytometry - Anti-CD40 antibody [EPR20735] (ab224639)](https://www.abcam.com/ps/products/224/ab224639/Images/ab224639-293849-anti-cd40-antibody-epr20735-flow-cytometry.jpg)
Flow Cytometry - Anti-CD40 antibody [EPR20735] (ab224639)Flow cytometric analysis of Raji (human Burkitt's lymphoma cell line) cell line labeling CD40 with ab224639 at 1/600 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077), at 1/2000 dilution was used as the secondary antibody.
Gated on total viable cells.
-
![Immunocytochemistry/ Immunofluorescence - Anti-CD40 antibody [EPR20735] (ab224639)](https://www.abcam.com/ps/products/224/ab224639/Images/ab224639-293848-anti-cd40-antibody-epr20735-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-CD40 antibody [EPR20735] (ab224639)Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Raji (human Burkitt's lymphoma cell line) cells labeling CD40 with ab224639 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous and weakly cytoplasmic staining in Raji cell line.
Negative control: Jurkat T cell line (PMID:10498643).
The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).
Secondary antibody only control: PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
-
![Western blot - Anti-CD40 antibody [EPR20735] (ab224639)](https://www.abcam.com/ps/products/224/ab224639/Images/ab224639-293846-anti-cd40-antibody-epr20735-western-blot.jpg)
Western blot - Anti-CD40 antibody [EPR20735] (ab224639)All lanes : Anti-CD40 antibody [EPR20735] (ab224639) at 1/1000 dilution
Lane 1 : Human lymph node tissue lysate
Lane 2 : Human lymphoma tissue lysate
Lane 3 : Raji (human Burkitt's lymphoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Developed using the ECL technique.
Predicted band size: 30 kDa
Observed band size: 42 kDa why is the actual band size different from the predicted?
Exposure time: 15 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
-
![Immunoprecipitation - Anti-CD40 antibody [EPR20735] (ab224639)](https://www.abcam.com/ps/products/224/ab224639/Images/ab224639-293850-anti-cd40-antibody-epr20735-immunoprecipitation.jpg)
Immunoprecipitation - Anti-CD40 antibody [EPR20735] (ab224639)CD40 was immunoprecipitated from 0.35 mg of Raji (human Burkitt's lymphoma cell line) whole cell lysate with ab224639 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab224639 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.
Lane 1: Raji whole cell lysate 10 μg (Input).
Lane 2: ab224639 IP in Raji whole cell lysate (+).
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab224639 in Raji whole cell lysate (-).Exposure time: 3 minutes.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR20735] (ab224639)](https://www.abcam.com/ps/products/224/ab224639/Images/ab224639-293853-anti-cd40-antibody-epr20735-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR20735] (ab224639)Immunohistochemical analysis of paraffin-embedded human diffuse large B-cell lymphoma tissue labeling CD40 with ab224639 at 1/250 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous staining in human diffuse large B-cell lymphoma tissue (PMID: 20593977) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR20735] (ab224639)](https://www.abcam.com/ps/products/224/ab224639/Images/ab224639-293854-anti-cd40-antibody-epr20735-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR20735] (ab224639)Immunohistochemical analysis of paraffin-embedded human Hodgkin's lymphoma tissue labeling CD40 with ab224639 at 1/250 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Membranous and cytoplasmic staining in Reed-Sternberg cells and tumor cells of human Hodgkin's lymphoma (PMID:7507299) is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
-
![Anti-CD40 antibody [EPR20735] (ab224639)](https://www.abcam.com/ps/products/224/ab224639/Images/ab224639-9-benefits-of-recombinant-antibodies.png)
Anti-CD40 antibody [EPR20735] (ab224639)
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
-
SDS download
-
Datasheet download
Certificate of Compliance
References (6)
ab224639 has been referenced in 6 publications.
- Luo C et al. CD40 is Positively Correlated with the Expression of Nucleophosmin in Cisplatin-Resistant Bladder Cancer. J Oncol 2020:3676751 (2020). PubMed: 32411234
- Zhou ZQ et al. Adipose extracellular matrix promotes skin wound healing by inducing the differentiation of adipose-derived stem cells into fibroblasts. Int J Mol Med 43:890-900 (2019). PubMed: 30535488
- Wang L et al. Identification of circular RNA Hsa_circ_0001879 and Hsa_circ_0004104 as novel biomarkers for coronary artery disease. Atherosclerosis 286:88-96 (2019). PubMed: 31103880
- Jiang C et al. CRISPR/Cas9 Screens Reveal Multiple Layers of B cell CD40 Regulation. Cell Rep 28:1307-1322.e8 (2019). PubMed: 31365872
- Hong D et al. Human-induced pluripotent stem cell-derived macrophages and their immunological function in response to tuberculosis infection. Stem Cell Res Ther 9:49 (2018). WB . PubMed: 29482598
- Journo G et al. Modulation of Cellular CpG DNA Methylation by Kaposi's Sarcoma-Associated Herpesvirus. J Virol 92:N/A (2018). PubMed: 29899086