Anti-CD3抗体[SP7] (ab16669)
Key features and details
- Rabbit monoclonal [SP7] to CD3
- Suitable for: Flow Cyt, IHC-P, WB, mIHC
- Reacts with: Mouse, Rat, Human
- Isotype: IgG
概述
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产品名称
Anti-CD3抗体[SP7]
参阅全部 CD3 一抗 -
描述
兔单克隆抗体[SP7] to CD3 -
宿主
Rabbit -
Tested Applications & Species
Application Species Flow Cyt HumanIHC-P MouseRatHumanmIHC HumanWB MouseRatHuman -
免疫原
Synthetic peptide within Human CD3 aa 150 to the C-terminus. The exact sequence is proprietary.
Database link: P07766 -
阳性对照
- WB: Recombinant Human CD3 epsilon protein (ab114153), Jurkat whole cell lysate. Human, mouse and rat thymus tissue lysate. IHC-P: Rat spleen tissue. Human tonsil tissue. Mouse epidydimal fat pad and lymph node tissues. Rat infarcted heart and spleen tissues. Flow Cyt: Human peripheral blood lymphocytes. Jurkat cells.
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常规说明
FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.
We recommend ab135372 as an alternative.
This antibody is suitable for staining normal and neoplastic T cells in formalin-fixed, paraffin-embedded tissues.
Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).
See other anti-rabbit secondary antibodies that can be used with this antibody.
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.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid repeated freeze / thaw cycles. -
存储溶液
pH: 7.50
Preservative: 0.1% Sodium azide
Constituents: Tissue culture supernatant, Tris buffered saline, 1% BSA -
Concentration information loading...
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纯度
Tissue culture supernatant -
Primary antibody说明
This antibody is suitable for staining normal and neoplastic T cells in formalin-fixed, paraffin-embedded tissues. -
克隆
单克隆 -
克隆编号
SP7 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab16669 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
应用 | Species |
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Flow Cyt |
Human
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IHC-P |
Mouse
Rat
Human
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mIHC |
Human
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WB |
Mouse
Rat
Human
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All applications |
Sheep
Rabbit
Horse
Chicken
Cow
Cat
Dog
Pig
Baboon
Woodchuck
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应用 | Ab评论 | 说明 |
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Flow Cyt | (2) | |
IHC-P | (51) |
1/150. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Boil tissue section in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at room temperature for 20 min. |
WB | (1) |
1/25. Predicted molecular weight: 23 kDa.
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mIHC |
Use at an assay dependent concentration.
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说明 |
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IHC-P
1/150. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. Boil tissue section in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at room temperature for 20 min. |
WB
1/25. Predicted molecular weight: 23 kDa. |
mIHC
Use at an assay dependent concentration. |
靶标
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功能
The CD3 complex mediates signal transduction. -
疾病相关
Defects in CD3D are a cause of severe combined immunodeficiency autosomal recessive T-cell-negative/B-cell-positive/NK-cell-positive (T(-)/B(+)/NK(+) SCID) [MIM:608971]. A form of severe combined immunodeficiency (SCID), a genetically and clinically heterogeneous group of rare congenital disorders characterized by impairment of both humoral and cell-mediated immunity, leukopenia, and low or absent antibody levels. Patients present in infancy recurrent, persistent infections by opportunistic organisms. The common characteristic of all types of SCID is absence of T-cell-mediated cellular immunity due to a defect in T-cell development. -
序列相似性
Contains 1 ITAM domain. -
细胞定位
Membrane. - Information by UniProt
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数据库链接
- Entrez Gene: 915 Human
- Entrez Gene: 916 Human
- Entrez Gene: 917 Human
- Entrez Gene: 919 Human
- Entrez Gene: 12500 Mouse
- Entrez Gene: 12501 Mouse
- Entrez Gene: 12502 Mouse
- Entrez Gene: 12503 Mouse
see all -
别名
- 4930549J05Rik antibody
- A430104F18Rik antibody
- AW552088 antibody
see all
图片
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 antibody [SP7] (ab16669)
IHC image of CD3 staining in a formalin fixed, paraffin embedded normal rat spleen tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab16669 at 1/100 dilution for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 antibody [SP7] (ab16669)
IHC image of CD3 staining in a formalin fixed, paraffin embedded normal human tonsil tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab16669 at 1/100 dilution for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 antibody [SP7] (ab16669)
IHC image of CD3 staining in mouse lymph node formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol B. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab16669 at 1/100 dilution for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times
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Human peripheral blood lymphocytes stained with ab16669 (red line). Human whole blood was processed using a modified protocol based on Chow et al, 2005 (PMID: 16080188). In brief, human whole blood was fixed in 4% formaldehyde (methanol-free) for 10 min at 22°C. Red blood cells were then lyzed by the addition of Triton X-100 (final concentration - 0.1%) for 15 min at 37°C. For experimentation, cells were treated with 50% methanol (-20°C) for 15 min at 4°C. Cells were then incubated with the antibody (ab16669, 1/1000 dilution) for 30 min at 4°C. The secondary antibody used was Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/2000 dilution for 30 min at 4°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >30,000 total events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. Gating strategy - peripheral blood lymphocytes.
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Fluorescence multiplex immunohistochemical analysis of normal human tonsil tissue (formalin-fixed paraffin-embedded section).
Merged staining of anti-PD1 (ab237728; orange; Opal™520), anti-PDL1 (ab237726; green; Opal™540), anti-CD68 (ab192847; yellow; Opal™570), anti-CD3 (ab16669; red; Opal™620), anti-Ki67 (ab16667; light blue; Opal™650) and anti-PanCK (ab7753; grey; Opal™690).
The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 7-color automation IHC kit (NEL821001KT, Akoya Biosciences®).
The section was incubated in six rounds of staining; in the order of ab237728 (1/500 dilution), ab237726 (1/500 dilution), ab192847 (1/300 dilution), ab16669 (1/300 dilution), ab16667 (1/200 dilution) and ab7753 (1/200 dilution); each using a separate fluorescent tyramide signal amplification system.
Sodium citrate antigen retrieval (Leica ER1, pH6.0, 30 minutes) was used in between rounds of tyramide signal amplification to remove the antibody from the previous round, to avoid any cross-reactivity.
DAPI (dark blue) was used as a nuclear counter stain.
Microscopy and pseudocoloring of individual Opal™ dyes was performed using a Vectra Polaris.
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All lanes : Anti-CD3 antibody [SP7] (ab16669) at 1/25 dilution
Lane 1 : THP1 whole cell lysate (-ve control)
Lane 2 : Raji whole cell lysate (-ve control)
Lane 3 : Jurkat whole cell lysate
Lane 4 : Human Thymus tissue lysate
Lane 5 : Mouse Thymus tissue lysate
Lane 6 : Rat Thymus tissue lysate
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 23 kDa
Observed band size: 23 kDaLanes 1 - 6: Merged signal (red and green). Green – ab16669 observed at 23 kDa. Red - loading control, ab8245, observed at 37 kDa.
This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using Licor blocking buffer before being incubated with ab16669 and ab8245 (loading control) overnight at 4°C. Antibody binding was detected using Goat Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) at a 1:10000 dilution for 1hr at room temperature and then imaged.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 antibody [SP7] (ab16669)Image from Graff JN et al.,Oncotarget 7(33), 52810 - 52817. Fig 2,; doi: 10.18632/oncotarget.10547. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/3.0/.
IHC using multi-spectral imaging on human lymph node (A-C) obtained from men with mCRPC. A) H+E staining and B) single-color images (plus nuclear stain; DAPI) of CD3 (ab16669), CD8 (ab101500), CD163, PD-L1, cytokeratin (CK), DAPI and C) merged. H+E stating at 20X magnification; multi-spectral images 200X magnification.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 antibody [SP7] (ab16669)This image is courtesy of an Abreview submitted by Carl Hobbs
Immunohistochemical analysis of Formaldehyde fixed, paraffin-embedded rat spleen tissue sections labelling CD3 with ab16669 at a dilution of 1/100. Biotin conjugated Goat Anti-Rabbit IgG at 1/300 dilution was used as the secondary antibody. Antigen retrieval was heat mediated using citric acid.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 antibody [SP7] (ab16669)This image is courtesy of an Abreview submitted by Ying Li.
Ab16669 staining CD3 in Mouse Epidydimal fat pad tissue sections by Immunohistochemistry (Formalin/PFA-fixed paraffin embedded sections). Tissue sections were fixed with formaldehyde, blocked with 5% serum for 4 hours at 25°C and permeabilized with Triton X-100. Samples were incubated with primary antibody (1/100 in PBST with BSA and goat serum) for 4°C at 12 hours. An Alexa Fluor® 568 goat anti-rabbit IgG (H + L) cross adsorbed was used as the secondary antibody.
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Flow cytometric analysis of rabbit anti-CD3 (SP7) antibody ab16669 (1/100) in Jurkats cells (green) compare to negative control of rabbit IgG (blue).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 antibody [SP7] (ab16669)This image is courtesy of an Abreview submitted by Dr Mal Niladri
ab16669 staining rat infarcted heart tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections).
Myocardial infarction was produced in a rat model following the ligation of the left anterior descending (LAD) coronary artery. Tissue was harvested 6 w following infarct, fixed with Histochoice for 72 hr, paraffin sectioned and the slide was then baked prior to CD3 staining. ab16669 at 1/200 was incubated overnight at 4°C. The image was taken with a confocal laser scanning microscope and shows cells giving strong inmmunofluorescence staining for CD3 antigen (green), indicating presence of cells of T-lymphocytes origin in the infarct zone of the heart tissue, counterstained nuclei with DAPI (blue). Note, CD3 tended to be present in nests of 2-5 cells that were non-uniformly distributed in the infarct zone. In addition, the image shows that the CD3 localization is predominantly membrane based and to a certain extent intracytoplasmic.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD3 antibody [SP7] (ab16669)Image from Moussion C et al., PLoS One. 2008 Oct 6;3(10):e3331. Fig 2.; doi:10.1371/journal.pone.0003331; October 6, 2008, PLoS ONE 3(10): e3331.Immunohistochemical analysis of Human tonsil tissue, staining CD3 (green) with ab16669.
Antigen retrieval was performed by heat mediation in citrate buffer (pH 6) and blocked with 5% goat serum and 5% BSA for 1 hour at room temperature. Samples were incubated with primary antibody (1/100) overnight at 4°C. A Cy3®-conjugated anti-rabbit IgG was used as the secondary antibody.
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (289)
ab16669 被引用在 289 文献中.
- Ando M et al. Long-term eradication of extranodal natural killer/T-cell lymphoma, nasal type, by induced pluripotent stem cell-derived Epstein-Barr virus-specific rejuvenated T cells in vivo. Haematologica 105:796-807 (2020). PubMed: 31296577
- Crespo-Moral M et al. Histological and immunohistochemical characterization of the porcine ocular surface. PLoS One 15:e0227732 (2020). PubMed: 31929592
- Runge EM et al. CD4+ T cell expression of the IL-10 receptor is necessary for facial motoneuron survival after axotomy. J Neuroinflammation 17:121 (2020). PubMed: 32303238
- Pietka W et al. Lack of interleukin-33 and its receptor does not prevent calcipotriol-induced atopic dermatitis-like inflammation in mice. Sci Rep 10:6451 (2020). PubMed: 32296080
- Chen T et al. Synthetic and immunological studies on the OCT4 immunodominant motif antigen-based anti-cancer vaccine. Cancer Biol Med 17:132-141 (2020). PubMed: 32296581