Anti-CD3抗体[CD3-12] (ab11089)
Key features and details
- Rat monoclonal [CD3-12] to CD3
- Suitable for: IHC-P, WB, Flow Cyt (Intra)
- Reacts with: Mouse, Rat, Human
- Isotype: IgG1
Related conjugates and formulations
概述
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产品名称
Anti-CD3抗体[CD3-12]
参阅全部 CD3 一抗 -
描述
大鼠单克隆抗体[CD3-12] to CD3 -
宿主
Rat -
经测试应用
适用于: IHC-P, WB, Flow Cyt (Intra)more details -
种属反应性
与反应: Mouse, Rat, Human
预测可用于: Sheep, Rabbit, Horse, Chicken, Cow, Cat, Dog, Pig, Duck, Cynomolgus monkey, Rhesus monkey, Woodchuck, Raccoon, Alpaca -
免疫原
Synthetic peptide corresponding to Human CD3. Derived from cytoplasmic epitope.
Database link: P07766 -
阳性对照
- IHC-P: Human tonsil tissue. Mouse spleen and lymph node tissue; Rat spleen tissue. Flow Cyt (Intra): Human peripheral blood lymphocytes. WB: Jurkat, MOLT-4 and EL4 cell lysates; Human, mouse and rat thymus tissue lysates.
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常规说明
This product should be stored undiluted. Storage in frost free freezers is not recommended. Should this product contain a precipitate we recommend microcentrifugation before use.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 this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As
性能
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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 or -80°C. Avoid freeze / thaw cycle. -
存储溶液
Preservative: 0.09% Sodium azide
Constituent: PBS -
Concentration information loading...
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纯度
Protein G purified -
纯化说明
Purified IgG prepared from tissue culture supernatant. -
克隆
单克隆 -
克隆编号
CD3-12 -
同种型
IgG1 -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab11089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 |
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IHC-P | (14) |
1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
1mM EDTA pH8.0 is recommended for this purpose. |
WB |
Use at an assay dependent concentration. Predicted molecular weight: 23.1 kDa.
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Flow Cyt (Intra) |
1/50 - 1/100.
Membrane permeabilization is recommended for this application. ab18407 - Rat monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
说明 |
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IHC-P
1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. 1mM EDTA pH8.0 is recommended for this purpose. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 23.1 kDa. |
Flow Cyt (Intra)
1/50 - 1/100. Membrane permeabilization is recommended for this application. ab18407 - Rat monoclonal IgG1, is suitable for use as an isotype control with this antibody. |
图片
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 0.1% Tween-20 permeabilized human peripheral blood mononuclear cells (PBMC) labeling CD3 with ab11089 at 1/400 dilution (red) compared with a Mouse IgG, monoclonal Isotype Control (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Mouse IgG ab150157 (Alexa Fluor®488) at 1/2000 dilution was used as the secondary antibody.
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All lanes : Anti-CD3 antibody [CD3-12] (ab11089) at 1 µg/ml
Lane 1 : THP-1 (Human monocytic leukemia cell line) whole cell lysate (negative control)
Lane 2 : Raji (Human Burkitt's lymphoma cell line) whole cell lysate (negative control)
Lane 3 : Jurkat (Human T cell leukemia cell line from peripheral blood) 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-Rat at 1/10000 dilution
Performed under reducing conditions.
Predicted band size: 23.1 kDa
Observed band size: 23 kDa why is the actual band size different from the predicted?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 ab11089 overnight at 4°C. Antibody binding was detected using Goat anti-Rat secondary at a 1:10000 dilution for 1hr at room temperature and then imaged.
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Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling CB3 with ab11089 at 1/200 dilution, followed by ready to use Goat Anti-rat IgG H&L (HRP polymer) (ab214882). Positive staining on rat spleen tissue is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, followed by ready to use Goat Anti-Mouse IgG H&L (HRP polymer) (ab214882). Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD3 with ab11089 at 1/200 dilution, followed by ready to use Goat Anti-Rat IgG H&L (HRP polymer) (ab214882). Positive staining on mouse spleen tissue is observed. Counterstained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, followed by ready to use Goat Anti-Mouse IgG H&L (HRP polymer) (ab214882). Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).
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IHC image of CD3 staining in a formalin fixed, paraffin embedded human tonsil tissue. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH 6). The section was incubated with ab11089 at 1/250 dilution for 15 minutes at room temperature. A goat anti-rat biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. The section was 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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IHC image of CD3 staining in a formalin fixed, paraffin embedded normal mouse lymph node tissue.
The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH 6). The section was incubated with ab11089 at 1/250 dilution for 15 minutes at room temperature. A goat anti-rat biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. The section was 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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ab11089 staining CD3 in mouse lymph node tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections).
Tissue was fixed with formaldehyde and blocked with 15% serum for 60 minutes at 20°C. Antigen retrieval was by heat mediation in sodium citrate (pH 6). Samples were incubated with primary antibody (1/250 in TBS) for 18 hours at 20°C. An Alexa Fluor®conjugated goat anti-rat IgG polyclonal (1/400) was used as the secondary antibody.
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All lanes : Anti-CD3 antibody [CD3-12] (ab11089) at 1/1000 dilution
Lane 1 : Mouse thymus tissue lysate
Lane 2 : Rat thymus tissue lysate
Lane 3 : Human thymus tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution
Predicted band size: 23.1 kDaBlocking and dilution buffer: 5% NFDM/TBST.
Exposure: Lanes 1/3: 5 secs; Lanes 2: 3 secs.
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All lanes : Anti-CD3 antibody [CD3-12] (ab11089) at 1/1000 dilution
Lane 1 : Jurkat (human T cell leukemia T lymphocyte)
Lane 2 : MOLT-4 (human lymphoblastic leukemia T lymphoblast)
Lane 3 : EL4 (mouse lymphoma T lymphocyte) whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rat IgG H&L (HRP) (ab205720) at 1/5000 dilution
Predicted band size: 23.1 kDaBlocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 5 secs.
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Anti-CD3 antibody [CD3-12] (ab11089) + Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Secondary
HRP-conjugated goat anti-rat IgG
Predicted band size: 23.1 kDa
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (85)
ab11089 被引用在 85 文献中.
- Song Z et al. Integrated histopathological, lipidomic, and metabolomic profiles reveal mink is a useful animal model to mimic the pathogenicity of severe COVID-19 patients. Signal Transduct Target Ther 7:29 (2022). PubMed: 35091528
- Meschkat M et al. White matter integrity in mice requires continuous myelin synthesis at the inner tongue. Nat Commun 13:1163 (2022). PubMed: 35246535
- West AJ et al. Inflammasome-Associated Gastric Tumorigenesis Is Independent of the NLRP3 Pattern Recognition Receptor. Front Oncol 12:830350 (2022). PubMed: 35299732
- Yang J et al. Dynamic profiling of immune microenvironment during pancreatic cancer development suggests early intervention and combination strategy of immunotherapy. EBioMedicine 78:103958 (2022). PubMed: 35316682
- Nunez N et al. The unforeseen intracellular lifestyle of Enterococcus faecalis in hepatocytes. Gut Microbes 14:2058851 (2022). PubMed: 35373699