CD40重组抗体[RM1193]_CD40抗体(ab317587)| Abcam中文官网

产品名称

Anti-CD40抗体[RM1193] - BSA and Azide free
参阅全部 CD40 一抗
描述

兔重组multiclonal [RM1193] to CD40 - BSA and Azide free
宿主

Rabbit
特异性

Unsuitable for mouse ICC and mouse IP.
经测试应用

适用于: IHC-P, IP, ICC/IF, Flow Cyt, WBmore details
种属反应性

与反应: Mouse, Human
不与反应: Rat
免疫原

This product was produced with the following immunogens:
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
阳性对照
- WB: Daudi whole cell, Raji whole cell, RAW 264.7 treated with 1µg/ml LPS whole cell, A20 whole cell, Human lymph node tissue, Mouse thymus tissue and Mouse spleen tissue lysates. IHC-P: Human tonsil, Human colon, Human diffuse large B- lymphoma, Human hodgkin's lymphoma, Mouse spleen and Mouse colon tissues. ICC/IF: Raji cell. Flow Cyt: Raji cell. IP: ab317586 IP in Raji cell.
常规说明
ab317587 is the carrier-free version of ab317586
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.
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.

形式

Liquid
存放说明

Shipped at 4°C. Store at +4°C.
存储溶液

pH: 7.2
Constituent: 100% PBS
无载体

是
Concentration information loading...
纯度

Protein A purified
克隆

Recombinant Multiclonal
克隆编号

RM1193
同种型

IgG
研究领域
- Neuroscience
- Processes

Alternative Versions
- Anti-CD40 antibody [RM1193] (ab317586)
Compatible Secondaries
- VeriBlot for IP Detection Reagent (HRP) (ab131366)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Related Products

The Abpromise guarantee

Abpromise™承诺保证使用ab317587于以下的经测试应用

“应用说明”部分下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用	Ab评论	说明
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.
IP		Use at an assay dependent concentration.
ICC/IF		Use at an assay dependent concentration.
Flow Cyt		Use at an assay dependent concentration.
WB		Use at an assay dependent concentration.

说明
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.
IP Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.
WB Use at an assay dependent concentration.

功能

Receptor for TNFSF5/CD40LG.
组织特异性

B-cells and in primary carcinomas.
疾病相关

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.
序列相似性

Contains 4 TNFR-Cys repeats.
细胞定位

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
数据库链接
- Entrez Gene: 958 Human
- Entrez Gene: 21939 Mouse
- Omim: 109535 Human
- SwissProt: P25942 Human
- SwissProt: P27512 Mouse
- Unigene: 472860 Human
- Unigene: 271833 Mouse
别名
- 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

Western blot - Anti-CD40 antibody [RM1193] - BSA and Azide free (ab317587)

All lanes : Anti-CD40 antibody [RM1193] (ab317586) at 1/1000 dilution

Lane 1 : Daudi (human Burkitt's lymphoma lymphoblast) whole cell lysate
Lane 2 : Raji (human Burkitts lymphoma B lymphocyte) whole cell lysate
Lane 3 : Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate
Lane 4 : RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
Lane 5 : RAW 264.7 treated with 1µg/ml LPS (Lipopolysaccharide) for 6 hours whole cell lysate
Lane 6 : A20 (mouse reticulum sarcoma B lymphocyte) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Observed band size: 42 kDa why is the actual band size different from the predicted?

This data was developed using ab317586, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Anti-CD40 antibody (ab317586) staining at 1/1000 dilution is specific to human and mouse species. Anti-CD40 antibody (ab13545) at 1/1000 dilution is bandy than ab317586.

The molecular weight observed is consistent with the literature (PMID: 23404288).

Negative control: Jurkat (PMID:10498643; PMID:15708600).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time: Lane 1, 6: 103 seconds Lane 2-5: 92 seconds
Western blot - Anti-CD40 antibody [RM1193] - BSA and Azide free (ab317587)

All lanes : Anti-CD40 antibody [RM1193] (ab317586) at 1/1000 dilution

Lane 1 : Human lymph node tissue lysate
Lane 2 : Mouse thymus tissue lysate
Lane 3 : Mouse spleen tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Developed using the ECL technique.

Observed band size: 42 kDa why is the actual band size different from the predicted?

Exposure time: 136 seconds

This data was developed using ab317586, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Anti-CD40 antibody (ab317586) staining at 1/1000 dilution is specific to human and mouse species. Anti-CD40 antibody (ab13545) at 1/1000 dilution is weak than ab317586 in mouse tissue lysate.

The molecular weight observed is consistent with the literature (PMID: 23404288).

The lysis conditions of the tissues could be further optimised to improve performance.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [RM1193] - BSA and Azide free (ab317587)

This data was developed using ab317586, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling CD40 with ab317586 at 1/2000 (0.254 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human tonsil in (A) ab317586 and (B) ab13545.
The image on (A) ab317586 is applied with Anti-CD40 antibody at 1/2000 dilution and (B) ab13545 is applied with Anti-CD40 antibody at 1/4000 dilution.
The section was incubated with ab317586 or ab13545 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [RM1193] - BSA and Azide free (ab317587)

This data was developed using ab317586, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling CD40 with ab317586 at 1/2000 (0.254 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on immune cells of human colon in (A) ab317586, non-specific staining on human colon in (B) ab13545. The image on (A) ab317586 is applied with Anti-CD40 antibody at 1/2000 dilution and (B) ab13545 is applied with Anti-CD40 antibody at 1/4000 dilution.
The section was incubated with ab317586 or ab13545 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [RM1193] - BSA and Azide free (ab317587)

This data was developed using ab317586, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human diffuse large B-cell lymphoma tissue labeling CD40 with ab317586 at 1/2000 (0.254 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human diffuse large B-cell lymphoma. The section was incubated with ab317586 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [RM1193] - BSA and Azide free (ab317587)

This data was developed using ab317586, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human hodgkin's lymphoma tissue labeling CD40 with ab317586 at 1/2000 (0.254 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human hodgkin's lymphoma. The section was incubated with ab317586 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [RM1193] - BSA and Azide free (ab317587)

This data was developed using ab317586, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling CD40 with ab317586 at 1/500 (1.016 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse spleen. The section was incubated with ab317586 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [RM1193] - BSA and Azide free (ab317587)

This data was developed using ab317586, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling CD40 with ab317586 at 1/500 (1.016 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse colon. The section was incubated with ab317586 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [RM1193] - BSA and Azide free (ab317587)

This data was developed using ab317586, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Human cardiac muscle tissue labeling CD40 with ab317586 at 1/2000 (0.254 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control: no staining on human cardiac muscle. The section was incubated with ab317586 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [RM1193] - BSA and Azide free (ab317587)

This data was developed using ab317586, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling CD40 with ab317586 at 1/500 (1.016 ug/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Negative control: no staining on mouse cardiac muscle. The section was incubated with ab317586 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins
Immunocytochemistry/ Immunofluorescence - Anti-CD40 antibody [RM1193] - BSA and Azide free (ab317587)

This data was developed using ab317586, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Raji (human Burkitt's lymphoma B lymphocyte) cells labelling CD40 with ab317586 at 1/50 (10.16 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green).

Confocal image showing membranous staining in Raji cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labeled with DAPI (shown in blue).
Negative control: Jurkat (PMID: 10498643, PMID: 15708600).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5 ug/ml dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.
Flow Cytometry - Anti-CD40 antibody [RM1193] - BSA and Azide free (ab317587)

This data was developed using ab317586, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of Jurkat(human T cell leukemia T lymphocyte from peripheral blood, Left) / Raji(human Burkitt's lymphoma B lymphocyte, Right) cells labelling CD40 with ab317586 at 1/500 dilution (0.1 ug)/Red compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Gated on viable cell.
Negative control: Jurkat.
Immunoprecipitation - Anti-CD40 antibody [RM1193] - BSA and Azide free (ab317587)

This data was developed using ab317586, the same antibody clone in a different buffer formulation.
CD40 was immunoprecipitated from 0.35 mg Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate with ab317586 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab317586 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate
Lane 2: ab317586 IP in Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab317586 in Raji whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 111 seconds.
Anti-CD40 antibody [RM1193] - BSA and Azide free (ab317587)

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 download

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概述

产品名称

描述

宿主

特异性

经测试应用

种属反应性

免疫原

阳性对照

常规说明

性能

形式

存放说明

存储溶液

无载体

纯度

克隆

克隆编号

同种型

研究领域

相关产品

Alternative Versions

Compatible Secondaries

Isotype control

Related Products

应用

The Abpromise guarantee

靶标

功能

组织特异性

疾病相关

序列相似性

细胞定位

数据库链接

别名

图片

实验方案

数据表及文件

Datasheet download

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