Recombinant Anti-CD40 antibody [EPR20540] - Low endotoxin, Azide free (ab223546)
![Mass Cytometry - Anti-CD40 antibody [EPR20540] - Low endotoxin, Azide free (ab223546)](https://www.abcam.com/ps/products/223/ab223546/Images/ab223546-355416-AntiCD40-antibody-EPR20540--Low-endotoxin-Azide-free-imagingmasscytometry-human-lung.png "Mass Cytometry - Anti-CD40 antibody [EPR20540] - Low endotoxin, Azide free (ab223546)")
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 - Low endotoxin, Azide free
- Suitable for: WB, IHC-P, Mass Cytometry
- Knockout validated
- Reacts with: Human
Overview
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Product name
Anti-CD40 antibody [EPR20540] - Low endotoxin, Azide free
See all CD40 primary antibodies -
Description
Rabbit monoclonal [EPR20540] to CD40 - Low endotoxin, Azide free -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, Mass Cytometrymore 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. IMC: Human tonsil tissue.
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General notes
ab223546 is the carrier-free version of ab213205.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
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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Conjugation kits
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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 ab223546 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 |
Use at an assay dependent concentration. Detects a band of approximately 42 kDa (predicted molecular weight: 30 kDa).
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| IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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| Mass Cytometry |
Use at an assay dependent concentration.
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| Notes |
|---|
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WB
Use at an assay dependent concentration. Detects a band of approximately 42 kDa (predicted molecular weight: 30 kDa). |
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IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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Mass Cytometry
Use at an assay dependent concentration. |
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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Mass Cytometry - Anti-CD40 antibody [EPR20540] - Low endotoxin, Azide free (ab223546)This image is courtesy of the Single Cell & Imaging Mass Cytometry Analysis Platform, Goodman Cancer Research Centre, McGill University
Imaging Mass Cytometry™ (IMC™) image of human tonsil tissue stained with Anti-CD40 antibody [EPR20540]. ab223546 (carrier-free antibody, purified) was metal-conjugated using a Maxpar® Antibody Labeling Kit from Fluidigm. Immunostaining was performed according to Fluidigm’s protocols. Briefly, slides were subject to deparaffinization and heat-induced epitope retrieval, followed by overnight incubation at 4°C with an antibody cocktail containing metal-tagged antibodies in blocking buffer. Slides were subsequently washed with 0.2% Triton-X and 1x PBS, counterstained with Cell-ID™ Intercalator-Ir diluted at 1/400 in 1x PBS for 30 min at room temperature, rinsed for 5 min with distilled H2O, and air-dried prior to IMC™ acquisition. IMC™ acquisition was performed using the Fluidigm Hyperion™ Imaging System.
Imaging Mass Cytometry™, IMC™, Cell-ID™, Hyperion™ and Maxpar® are trademarks of Fluidigm Canada
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![Western blot - Anti-CD40 antibody [EPR20540] - Low endotoxin, Azide free (ab223546)](https://www.abcam.com/ps/products/223/ab223546/Images/ab223546-380883-anti-cd40-antibody-epr20540-low-endotoxin-azide-free-western-blot-u2-os-wt-u2-os-cd40-knockout-raji-mcf7.jpg)
Western blot - Anti-CD40 antibody [EPR20540] - Low endotoxin, Azide free (ab223546)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?This data was developed using the same antibody clone in a different buffer formulation (ab213205).
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] - Low endotoxin, Azide free (ab223546)](https://www.abcam.com/ps/products/223/ab223546/Images/ab223546-325010-anti-cd40-antibody-epr20540-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR20540] - Low endotoxin, Azide free (ab223546)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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab213205).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR20540] - Low endotoxin, Azide free (ab223546)](https://www.abcam.com/ps/products/223/ab223546/Images/ab223546-282800-anti-cd40-antibody-epr20540-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR20540] - Low endotoxin, Azide free (ab223546)This IHC data was generated using the same anti-CD40 antibody clone [EPR20540] in a different buffer formulation (cat# 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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![Anti-CD40 antibody [EPR20540] - Low endotoxin, Azide free (ab223546)](https://www.abcam.com/ps/products/223/ab223546/Images/ab223546-5-benefits-of-recombinant-antibodies.png)
Anti-CD40 antibody [EPR20540] - Low endotoxin, Azide free (ab223546)
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (0)
ab223546 has not yet been referenced specifically in any publications.
