製品の概要

  • 製品名
    Anti-CD3 antibody [SP7]
    CD3 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [SP7] to CD3
  • アプリケーション
    適用あり: Flow Cyt, IHC-Fr, IHC-P, WB, IHC-FoFrmore details
  • 種交差性
    交差種: Mouse, Rat, Human, Pig
    交差が予測される動物種: Sheep, Rabbit, Horse, Chicken, Cow, Cat, Dog, Baboon, Woodchuck
  • 免疫原

    Synthetic peptide:

    KAKAKPVTRGAGA

    , corresponding to amino acids 156-168 of Human CD3 epsilon chain.

  • エピトープ
    aa 156-168 of epsilon chain of Human CD3 protein (intracytoplasmic).
  • ポジティブ・コントロール
    • Tonsil IHC: Pig and rat spleen tissue
  • 特記事項

    This antibody is suitable for staining normal and neoplastic T cells in formalin-fixed, paraffin-embedded tissues.

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
Flow Cyt 1/1000. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IHC-Fr Use at an assay dependent concentration. PubMed: 18658050
IHC-P 1/100. 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 1/200. Predicted molecular weight: 23 kDa.
IHC-FoFr Use at an assay dependent concentration.

ターゲット情報

  • 機能
    The CD3 complex mediates signal transduction.
  • 関連疾患
    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.
  • 配列類似性
    Contains 1 ITAM domain.
  • 細胞内局在
    Membrane.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 4930549J05Rik antibody
    • A430104F18Rik antibody
    • AW552088 antibody
    • Cd247 antibody
    • CD247 antigen antibody
    • CD247 antigen, zeta subunit antibody
    • CD247 molecule antibody
    • CD3 antibody
    • CD3 antigen, delta subunit antibody
    • CD3 delta antibody
    • CD3 epsilon antibody
    • CD3 eta antibody
    • CD3 gamma antibody
    • CD3 molecule delta polypeptide antibody
    • CD3 molecule, epsilon polypeptide antibody
    • CD3 molecule, gamma polypeptide antibody
    • CD3 zeta antibody
    • CD3-DELTA antibody
    • CD3d antibody
    • CD3D antigen delta polypeptide antibody
    • CD3d antigen, delta polypeptide (TiT3 complex) antibody
    • CD3d molecule delta antibody
    • CD3d molecule delta CD3 TCR complex antibody
    • CD3d molecule, delta (CD3-TCR complex) antibody
    • CD3D_HUMAN antibody
    • CD3E antibody
    • CD3e antigen antibody
    • CD3E antigen epsilon polypeptide antibody
    • CD3e antigen, epsilon polypeptide (TiT3 complex) antibody
    • CD3e molecule epsilon antibody
    • CD3e molecule epsilon CD3 TCR complex antibody
    • CD3e molecule, epsilon (CD3-TCR complex) antibody
    • CD3epsilon antibody
    • CD3G antibody
    • CD3g antigen antibody
    • CD3G antigen gamma polypeptide antibody
    • CD3g antigen, gamma polypeptide (TiT3 complex) antibody
    • CD3g molecule gamma antibody
    • CD3g molecule gamma CD3 TCR complex antibody
    • CD3g molecule, gamma (CD3-TCR complex) antibody
    • CD3H antibody
    • CD3Q antibody
    • CD3Z antibody
    • CD3zeta antibody
    • Ctg3 antibody
    • FLJ17620 antibody
    • FLJ17664 antibody
    • FLJ18683 antibody
    • FLJ79544 antibody
    • FLJ94613 antibody
    • IMD19 antibody
    • Leu-4 antibody
    • MGC138597 antibody
    • OKT3, delta chain antibody
    • OTTHUMP00000032544 antibody
    • T cell receptor antibody
    • T cell receptor T3 delta chain antibody
    • T cell receptor T3 gamma chain antibody
    • T cell receptor T3 zeta chain antibody
    • T cell receptor zeta chain antibody
    • T cell surface glycoprotein CD3 antibody
    • T cell surface glycoprotein CD3 delta chain antibody
    • T cell surface glycoprotein CD3 epsilon chain antibody
    • T cell surface glycoprotein CD3 gamma chain antibody
    • T cell surface glycoprotein CD3 zeta chain antibody
    • T-cell antigen receptor complex, delta subunit of T3 antibody
    • T-cell antigen receptor complex, epsilon subunit of T3 antibody
    • T-cell antigen receptor complex, gamma subunit of T3 antibody
    • T-cell antigen receptor complex, zeta subunit of CD3 antibody
    • T-cell receptor T3 delta chain antibody
    • T-cell receptor T3 gamma chain antibody
    • T-cell surface antigen T3/Leu-4 epsilon chain antibody
    • T-cell surface glycoprotein CD3 delta chain antibody
    • T-cell surface glycoprotein CD3 gamma chain antibody
    • T3 antibody
    • T3d antibody
    • T3e antibody
    • T3g antibody
    • T3z antibody
    • TCRE antibody
    • TCRk antibody
    • Tcrz antibody
    • TCRzeta antibody
    see all

画像

  • 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
    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 kDa

    Lanes 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 anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at a 1:10000 dilution for 1hr at room temperature and then imaged.

  • Immunohistochemical analysis of Formaldehyde fixed, paraffin-embedded pig spleen tissue sections labelling CD3 with ab16669 at a dilution of 1/100. Biotin conjugated Goat Anti-Rabbit IgG was used as the secondary antibody. Antigen retrieval was heat mediated using citric acid.

    See Abreview

  • Immunohistochemical (Formalin / PFA fixed paraffin-embedded sections) staining of of Tonsil tissue labeling CD3 (SP7) with ab16669 at 1/150 dilution.

  • 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.

    See Abreview

  • Immunohistochemical analysis of Formaldehyde fixed, frozen rat spleen tissue sections labelling CD3 with ab16669 at a dilution of 1/500. Biotin conjugated Goat Anti-Rabbit IgG at 1/300 dilution was used as the secondary antibody. 

    Cryostat sections (10 microns thick) of fresh frozen spleen were dried overnight. They were fixed in 10% Formalin in PBS pH7 for 15 minutes.
    After secondary antibody incubation ( Gt anti Rb -biotin) , stAvidin-Alexa 594 was applied.

    See Abreview

  • 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 Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) 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.

  • Flow cytometric analysis of rabbit anti-CD3 (SP7) antibody ab16669 (1/100) in Jurkats cells (green) compare to negative control of rabbit IgG (blue).

  • Human tonsil stained with ab16669.
  • 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.

    See Abreview

  • Anti-CD3 antibody [SP7] (ab16669) at 1/25 dilution + Jurkat cell lysate

    Predicted band size : 23 kDa
    Observed band size : 19 kDa (why is the actual band size different from the predicted?)
  • ab16669 staining CD3 in murine brain tissue by Immunohistochemistry (Frozen sections).
    Tissue was fixed in acetone, blocked using 5% serum for 30 minutes at 25°C and then incubated with ab16669 at a 1/200 dilution for 18 hours at 4°C. The secondary used was an undiluted HRP conjugated goat anti-rabbit polyclonal.

    See Abreview

  • 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.

参考文献

This product has been referenced in:
  • Amrouche L  et al. MicroRNA-146a in Human and Experimental Ischemic AKI: CXCL8-Dependent Mechanism of Action. J Am Soc Nephrol 28:479-493 (2017). Read more (PubMed: 27444565) »
  • Yu W  et al. A Systems Pharmacology Approach to Determine Active Compounds and Action Mechanisms of Xipayi KuiJie'an enema for Treatment of Ulcerative colitis. Sci Rep 7:1189 (2017). IHC-P ; Mouse . Read more (PubMed: 28446747) »

See all 81 Publications for this product

レビューと Q&A

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Sample
Dog Cell (skeletal muscle)
Specification
skeletal muscle
Permeabilization
No
Fixative
acetone/methanol 1:1 cold
Username

Dr. Ling Guo

Verified customer

投稿 Feb 10 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Dog Tissue sections (Tonsil)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate 6
Permeabilization
No
Specification
Tonsil
Blocking step
Protein Block Serum Free as blocking agent for 10 minute(s) · Concentration: 100% · Temperature: RT°C
Fixative
10% NBF
Username

Abcam user community

Verified customer

投稿 Jul 27 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Rat Tissue sections (liver)
Permeabilization
Yes - 0.05% Triton X-100
Specification
liver
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Jun 27 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rhesus monkey Tissue sections (Gastrointestinal tract)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris-EDTA/Ventana CC1
Permeabilization
No
Specification
Gastrointestinal tract
Fixative
Paraformaldehyde
Username

Lina Li

Verified customer

投稿 May 16 2017

can confirm that ab16669 CD3 antibody [SP7] is sold as tissue culture supernatant. Unpurified antibodies, such as those sold as whole antiserum, ascites or tissue culture supernatant will not have a concentration stated on the datasheet. Antibody conce...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 10mM Citrate
Permeabilization
No
Specification
Kidney
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Fixative
Formaldehyde
Username

Dr. Wesley Konsavage

Verified customer

投稿 Dec 26 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Chicken Tissue sections (Spleen)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Spleen
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

投稿 Dec 21 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Cow Tissue sections (Spleen)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Spleen
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

投稿 Dec 21 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Sheep Tissue sections (Lamb spleen)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Specification
Lamb spleen
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

投稿 Dec 21 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Common marmoset Tissue sections (Spleen)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Spleen
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2%
Fixative
Formaldehyde
Username

Mr. Carl Hobbs

Verified customer

投稿 Dec 21 2016

1-10 of 66 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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