CD27重组抗体[EPR8569]_CD27抗体(ab131254)| Abcam中国

产品名称

Anti-CD27抗体[EPR8569]
参阅全部 CD27 一抗
描述

兔单克隆抗体[EPR8569] to CD27
宿主

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Human
ICC/IF	Human
IHC-P	Human
WB	Human

See all applications and species data

免疫原

Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human CD27 aa 200 to the C-terminus (intracellular).
(Peptide available as ab192535)
阳性对照
- WB: Raji, Ramos and NAMALWA cell lysates and human lymph node and fetal spleen tissue lysates. IHC-P: Human stomach, lung carcinoma and tonsil tissues. ICC/IF: Jurkat cells. Flow Cyt: Ramos cells.
常规说明
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

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.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

Learn more here.

形式

Liquid
存放说明

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
解离常数（K_D）

K_D = 7.90 x 10 ^-11 M
Learn more about K_D
存储溶液

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 0.05% BSA, 40% Glycerol
Concentration information loading...
纯度

Protein A purified
克隆

单克隆
克隆编号

EPR8569
同种型

IgG
研究领域
- Cancer
- Tumor immunology
- Cytokines
- TNF

Alternative Versions
- Anti-CD27 antibody [EPR8569] - Low endotoxin, Azide free (ab192336)
- Anti-CD27 antibody [EPR8569] - BSA and Azide free (ab256583)
Compatible Secondaries
Conjugation kits
- PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
- APC Conjugation Kit - Lightning-Link® (ab201807)
Immunizing Peptide (Blocking)
- CD27 peptide (ab192535)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Positive Controls
- Ramos whole cell lysate (ab3955)
Recombinant Protein
- Recombinant Human CD27 protein (ab114342)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab131254 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

应用	Species
Flow Cyt	Human
ICC/IF	Human
IHC-P	Human
WB	Human

应用	Ab评论	说明
ICC/IF		1/250 - 1/500.
WB		1/1000. Detects a band of approximately 55 kDa (predicted molecular weight: 29 kDa).Can be blocked with CD27 peptide (ab192535). For unpurified use at 1/1000 - 1/10000.
IHC-P		1/1500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified use at 1/100 - 1/250.
Flow Cyt		1/100 - 1/500. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

说明
ICC/IF 1/250 - 1/500.
WB 1/1000. Detects a band of approximately 55 kDa (predicted molecular weight: 29 kDa).Can be blocked with CD27 peptide (ab192535). For unpurified use at 1/1000 - 1/10000.
IHC-P 1/1500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified use at 1/100 - 1/250.
Flow Cyt 1/100 - 1/500. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

功能

Receptor for CD70/CD27L. May play a role in survival of activated T-cells. May play a role in apoptosis through association with SIVA1.
组织特异性

Found in most T-lymphocytes.
序列相似性

Contains 3 TNFR-Cys repeats.
翻译后修饰

Phosphorylated and O-glycosylated.
细胞定位

Membrane.
Target information above from: UniProt accession P26842 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: 939 Human
- Omim: 186711 Human
- SwissProt: P26842 Human
- Unigene: 355307 Human
别名
- CD 27 antibody
- CD27 antibody
- CD27 antigen antibody
- CD27 molecule antibody
- CD27_HUMAN antibody
- CD27L receptor antibody
- LPFS2 antibody
- MGC20393 antibody
- OTTHUMP00000238557 antibody
- S152 antibody
- T cell activation antigen CD27 antibody
- T cell antivation antigen S152 antibody
- T-cell activation antigen CD27 antibody
- T14 antibody
- TNFRSF 7 antibody
- TNFRSF7 antibody
- TNFSF7 antibody
- Tp 55 antibody
- Tp55 antibody
- Tumor necrosis factor receptor superfamily member 7 antibody
see all

Flow Cytometry - Anti-CD27 antibody [EPR8569] (ab131254)

Flow Cytometry analysis of Ramos cells labelling CD27 with purified ab131254 at 1/300 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/150) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
Western blot - Anti-CD27 antibody [EPR8569] (ab131254)

All lanes : Anti-CD27 antibody [EPR8569] (ab131254) at 1/1000 dilution

Lane 1 : Wild-type Raji cell lysate
Lane 2 : CD27 knockout Raji cell lysate
Lane 3 : Ramos cell lysate
Lane 4 : Human brain tissue lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 29 kDa
Observed band size: 35 kDa why is the actual band size different from the predicted?

Lanes 1 - 4: Merged signal (red and green). Green - ab131254 observed at 35 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

ab131254 was shown to react with CD27 in wild-type Raji cells in western blot with loss of signal observed in CD27 knockout sample. Wild-type and CD27 knockout Raji cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween^®) before incubation with ab131254 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated 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.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD27 antibody [EPR8569] (ab131254)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD27 with purified ab131254 at 1/1500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500).

Negative control using PBS instead of primary antibody.

Counterstained with hematoxylin.
Western blot - Anti-CD27 antibody [EPR8569] (ab131254)

Anti-CD27 antibody [EPR8569] (ab131254) at 1/5000 dilution (purified) + NAMALWA (human Burkitt's lymphoma cell line) cell lysate at 20 µg

Secondary
Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

Predicted band size: 29 kDa
Observed band size: 55 kDa why is the actual band size different from the predicted?

Blocking and dilution buffer: 5% NFDM/TBST.
Western blot - Anti-CD27 antibody [EPR8569] (ab131254)

All lanes : Anti-CD27 antibody [EPR8569] (ab131254) at 1/1000 dilution (purified)

Lane 1 : Human lymph node tissue lysate
Lane 2 : Human fetal spleen tissue lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Peroxidase-conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution

Predicted band size: 29 kDa
Observed band size: 55 kDa why is the actual band size different from the predicted?

Blocking and dilution buffer: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD27 antibody [EPR8569] (ab131254)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue showing negative staining with purified ab131254 at 1/800.

Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500).

Negative control using PBS instead of primary antibody.

Counterstained with hematoxylin.
Immunocytochemistry/ Immunofluorescence - Anti-CD27 antibody [EPR8569] (ab131254)

Immunocytochemistry/Immunofluorescence analysis of Jurkat cells labelling CD27 with purified ab131254 at 1/500. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/500) were also used.
Control 1: primary antibody (1/500) and secondary antibody, ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/500).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500).
Western blot - Anti-CD27 antibody [EPR8569] (ab131254)

All lanes : Anti-CD27 antibody [EPR8569] (ab131254) at 1/1000 dilution (unpurified)

Lane 1 : Ramos (human Burkitt's lymphoma cell line) cell lysate
Lane 2 : Human lymph node cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : HRP labelled Goat anti Rabbit IgG at 1/2000 dilution

Predicted band size: 29 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD27 antibody [EPR8569] (ab131254)

Immunohistochemistry (Formalin/PFA-fixed paraffin embedded sections) analysis of human stomach tissue labelling CD27 with ab131254 at a dilution of 1/100.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD27 antibody [EPR8569] (ab131254)

Immunohistochemistry (Formalin/PFA-fixed paraffin embedded sections) analysis of human tonsil tissue labelling CD27 with ab131254 at a dilution of 1/100.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
OI-RD Scanning - Anti-CD27 antibody [EPR8569] (ab131254)

Equilibrium disassociation constant (K_D)
Learn more about K_D

Click here to learn more about K_D
Anti-CD27 antibody [EPR8569] (ab131254)

Flow cytometry protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

发表研究结果有使用 ab131254？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab131254 被引用在 10 文献中.

Ansell SM et al. Safety and activity of varlilumab, a novel and first-in-class agonist anti-CD27 antibody, for hematologic malignancies. Blood Adv 4:1917-1926 (2020). PubMed: 32380537
Dhaeze T et al. CD70 defines a subset of proinflammatory and CNS-pathogenic TH1/TH17 lymphocytes and is overexpressed in multiple sclerosis. Cell Mol Immunol 16:652-665 (2019). PubMed: 30635649
Inoue S et al. CD70 expression in tumor-associated fibroblasts predicts worse survival in colorectal cancer patients. Virchows Arch N/A:N/A (2019). PubMed: 30980190
Griss J et al. B cells sustain inflammation and predict response to immune checkpoint blockade in human melanoma. Nat Commun 10:4186 (2019). PubMed: 31519915
Gorris MAJ et al. Eight-Color Multiplex Immunohistochemistry for Simultaneous Detection of Multiple Immune Checkpoint Molecules within the Tumor Microenvironment. J Immunol 200:347-354 (2018). PubMed: 29141863
Pham-Ledard A et al. Molecular analysis of immunoglobulin variable genes supports a germinal center experienced normal counterpart in primary cutaneous diffuse large B-cell lymphoma, leg-type. J Dermatol Sci 88:238-246 (2017). PubMed: 28838616
Bolognesi MM et al. Multiplex Staining by Sequential Immunostaining and Antibody Removal on Routine Tissue Sections. J Histochem Cytochem 65:431-444 (2017). PubMed: 28692376
Wouters MC et al. Treatment Regimen, Surgical Outcome, and T-cell Differentiation Influence Prognostic Benefit of Tumor-Infiltrating Lymphocytes in High-Grade Serous Ovarian Cancer. Clin Cancer Res 22:714-24 (2016). PubMed: 26384738
Szyszko EA et al. Non-proliferating plasma cells detected in the salivary glands and bone marrow of autoimmune NOD.B10.H2b mice, a model for primary Sjögren's syndrome. Autoimmunity 49:41-9 (2016). PubMed: 26324998
Belone Ade F et al. Genome-Wide Screening of mRNA Expression in Leprosy Patients. Front Genet 6:334 (2015). IHC ; Human . PubMed: 26635870

Key features and details

概述

产品名称

描述

宿主

Tested Applications & Species

免疫原

阳性对照

常规说明

性能

形式

存放说明

解离常数（KD）

存储溶液

纯度

克隆

克隆编号

同种型

研究领域

相关产品

Alternative Versions

Compatible Secondaries

Conjugation kits

Immunizing Peptide (Blocking)

Isotype control

Positive Controls

Recombinant Protein

应用

The Abpromise guarantee

靶标

功能

组织特异性

序列相似性

翻译后修饰

细胞定位

数据库链接

别名

图片

实验方案

数据表及文件

SDS download

Datasheet download

文献 (10)

客户评价及客户问答

解离常数（K_D）