Anti-CD40 antibody [EPR20735] (ab224639)

Anti-CD40 antibody [EPR20735] (ab224639) is a rabbit monoclonal antibody detecting CD40 in Western Blot, Flow Cytometry, IP, IHC-P, ICC/IF. Suitable for Human.

- KO validated for confirmed specificity

- Biophysical QC for unrivalled batch-batch consistency

- Over 10 publications

Be the first to review this product! Submit a review

Related conjugates and formulations

ab228818
Carrier free
Anti-CD40 antibody [EPR20735] - BSA and Azide free
ab302971
Conjugated
Alexa Fluor® 488 Anti-CD40 antibody [EPR20735]
ab302800
Conjugated
Alexa Fluor® 594 Anti-CD40 antibody [EPR20735]
ab302830
Conjugated
Alexa Fluor® 647 Anti-CD40 antibody [EPR20735]
ab306538
Conjugated
APC Anti-CD40 antibody [EPR20735]
ab303603
Conjugated
PE Anti-CD40 antibody [EPR20735]
ab314949
Conjugated
FITC Anti-CD40 antibody [EPR20735]

Images

Flow Cytometry - Anti-CD40 antibody [EPR20735] (AB224639), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	Flow Cyt	WB	ICC/IF	IHC-P
Human	Tested	Tested	Tested	Tested	Tested

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/30	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/600	Notes Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/250	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Associated Products

Select an associated product type

15 products for Alternative Product

Target data

See full target information CD40

Function

Receptor for TNFSF5/CD40LG (PubMed:31331973). Transduces TRAF6- and MAP3K8-mediated signals that activate ERK in macrophages and B cells, leading to induction of immunoglobulin secretion (By similarity).

Alternative names

CD40, TNFRSF5, Tumor necrosis factor receptor superfamily member 5, B-cell surface antigen CD40, Bp50, CD40L receptor, CDw40

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR20735
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

What is this antibody validated in?

Anti-CD40 antibody [EPR20735] (ab224639) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Flow Cytometry (Flow Cyt), Immunoprecipitation (IP), Immunohistochemistry (IHC-P), Immunocytochemistry/immunofluorescence (ICC/IF) in Human samples.

What is the molecular weight of CD40?

Anti-CD40 [EPR20735] (ab224639) specifically detects a band for CD40 (UniProt: P25942) at a molecular weight of 30kDa.

Trusted by the scientific community

Anti-CD40 [EPR20735] (ab224639) was first used in a scientific publication in 2017 and has been cited over 10 times in peer-reviewed journals.

Trial sizes available!

Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies.

Specificity confirmed

The specificity of Anti-CD40 antibody [EPR20735] (ab224639) has been confirmed by Immunocytochemistry testing in CD40 Knockout U2OS cells.

Other related products

We have a range of other formats of antibody clone [EPR20735] also available for your convenience:
ab224639, Carrier free - ab228818, ab284427, Alexa Fluor® 594 - ab302800, Alexa Fluor® 647 - ab302830, Alexa Fluor® 488 - ab302971, PE - ab303603, FITC - ab314949

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

10 product images

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 (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) (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.
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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).
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 Anti-alpha Tubulin antibody [DM1A] - Loading Control 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) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed 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).
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) (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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab224639 in Raji whole cell lysate (-).
Exposure time: 3 minutes.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-CD40 antibody [EPR20735] (ab224639)
Developed using the ECL technique.
Predicted band size: 30 kDa
Observed band size: 42 kDa
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) (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) (Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker 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) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.
Western blot - Anti-CD40 antibody [EPR20735] (ab224639)
Blocking/Dilution buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-CD40 antibody [EPR20735] (ab224639) at 1/1000 dilution
Lane 1: Human lymph node tissue lysate at 20 µg
Lane 2: Human lymphoma tissue lysate at 20 µg
Lane 3: Raji (human Burkitt's lymphoma cell line) whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (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
Exposure time: 15s
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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).
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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0).
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR20735] (ab224639)
Tissue Microarrays stained for "Anti-CD40 antibody [EPR20735]" using "ab224639"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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). The sections were incubated with ab224639 at +4°C overnight followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).
Flow Cytometry - Anti-CD40 antibody [EPR20735] (ab224639)
Flow cytometry staining of human peripheral blood mononuclear cells (PBMCs) with ab224639 (right) or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (left). PBMCs were incubated for 30 min on ice in 1x PBS containing 10 µg/ml human IgG and 10 % normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab224639 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 106 in 100 µl at 0.2 μg/ml (1/10550)) for 30min on ice. The cells were simultaneously stained with CD19.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min on ice

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on viable cells.