Anti-CD40 抗体 [EPR20735] (ab224639)

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Immunocytochemistry/ Immunofluorescence - 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

製品名

Anti-CD40 antibody [EPR20735]
CD40 一次抗体製品一覧
製品の詳細

Rabbit monoclonal [EPR20735] to CD40
由来種

Rabbit
アプリケーション

適用あり: WB, IHC-P, Flow Cyt, IP, ICC/IFmore details
種交差性

交差種: Human
免疫原

Recombinant fragment within Human CD40 aa 1-200. The exact sequence is proprietary.
Database link: P25942
ポジティブ･コントロール
- 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)
特記事項
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
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

製品の状態

Liquid
保存方法

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.
バッファー

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
精製度

Protein A purified
ポリ/モノ

モノクローナル
クローン名

EPR20735
アイソタイプ

IgG
研究分野

Alternative Versions
- Anti-CD40 antibody [EPR20735] - BSA and Azide free (ab228818)
Compatible Secondaries
Conjugation kits
- PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
- APC Conjugation Kit - Lightning-Link® (ab201807)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant human CD40 protein (Active) (ab155710)
Related Products
- Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (ab195889)
- Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

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.

アプリケーション	Abreviews	特記事項
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.

機能

Receptor for TNFSF5/CD40LG.
組織特異性

B-cells and in primary carcinomas.
関連疾患

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.
配列類似性

Contains 4 TNFR-Cys repeats.
細胞内局在

Secreted and Cell membrane.
Target information above from: UniProt accession P25942 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
参照データベース
- Entrez Gene: 958 Human
- Omim: 109535 Human
- SwissProt: P25942 Human
- Unigene: 472860 Human
別名
- AI326936 antibody
- B cell associated molecule CD40 antibody
- B cell surface antigen CD40 antibody
- B cell-associated molecule antibody
- B-cell surface antigen CD40 antibody
- Bp50 antibody
- CD 40 antibody
- CD40 antibody
- CD40 antigen (TNF receptor superfamily member 5) antibody
- CD40 antigen antibody
- CD40 molecule antibody
- CD40 molecule, TNF receptor superfamily member 5 antibody
- CD40 protein antibody
- CD40 type II isoform antibody
- CD40L receptor antibody
- CDw40 antibody
- GP39 antibody
- HIGM1 antibody
- IGM antibody
- IMD3 antibody
- MGC9013 antibody
- Nerve growth factor receptor related B lymphocyte activation molecule antibody
- OTTHUMP00000031699 antibody
- OTTHUMP00000031700 antibody
- p50 antibody
- T-BAM antibody
- TBAM antibody
- TNF receptor superfamily member 5 antibody
- TNFRSF5 antibody
- TNR5_HUMAN antibody
- TRAP antibody
- Tumor necrosis factor receptor superfamily , member 5 antibody
- Tumor necrosis factor receptor superfamily member 5 antibody
- Tumor necrosis factor receptor superfamily member 5 precursor antibody
- Tumor necrosis factor receptor superfamily, member 5, isoform CRA_a antibody
see all

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)

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.
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)

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)

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 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
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)

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)

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)

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.

Click here to view the general protocols

Datasheet

ab224639 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab224639 は 6 報の論文で使用されています。

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

レビューと Q&A

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Key features and details

製品の概要

製品名

製品の詳細

由来種

アプリケーション

種交差性

免疫原

ポジティブ･コントロール

特記事項

製品の特性

製品の状態

保存方法

バッファー

精製度

ポリ/モノ

クローン名

アイソタイプ

研究分野

関連製品

Alternative Versions

Compatible Secondaries

Conjugation kits

Isotype control

Recombinant Protein

Related Products

アプリケーション

ターゲット情報

機能

組織特異性

関連疾患

配列類似性

細胞内局在

参照データベース

別名

画像

プロトコール

データシートおよび資料

Certificate of Compliance

参考文献 (6)

レビューと Q&A