製品の概要

  • 製品名
  • 製品の詳細
    Rabbit polyclonal to CD3
  • アプリケーション
    適用あり: IHC-Fr, WB, IHC-P, IHC-FoFr, ICC/IFmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide:

    KAKAKPVTRGAGA

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

  • ポジティブ・コントロール
    • WB: Jurkat cell lysate and rat thymus tissue lysate. IHC-P: Human tonsil, human spleen and mouse spleen tissues.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab5690 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
IHC-Fr 1/100. PubMed: 18628996
WB Use a concentration of 0.5 - 2 µg/ml. Predicted molecular weight: 23 kDa.
IHC-P 1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. The recommended starting incubation time is 10min.
IHC-FoFr Use at an assay dependent concentration. PubMed: 20126467
ICC/IF Use at an assay dependent concentration. PubMed: 23874197

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 (ab5690) at 1 µg/ml

    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 – ab5690 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 ab5690 and ab8245 (loading control) overnight at 4°C. Antibody binding was detected using 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.

  • 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) for 20 mins. The section was then incubated with ab5690 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

  • Lanes 1 - 2 : Anti-CD3 antibody (ab5690) at 1 µg/ml
    Lanes 3 - 4 : No primary antibody

    Lane 1 : Jurkat cell lysate at 30 µg
    Lane 2 : Rat thymus tissue lysate at 20 µg
    Lane 3 : Jurkat cell lysate at 30 µg
    Lane 4 : Rat thymus tissue lysate at 20 µg

    Secondary
    Goat anti-rabbit IgG (H+L), highly cross - adsorbed, HiLyte™ Fluor 750-labeled at 1/12500 dilution

    Predicted band size : 23 kDa
    Observed band size : 23 kDa
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labelling CD3 with ab5690 at 2µl/ml. Slides were steamed in IHC epitope retrieval solution for 35 minutes and then cooled for 20 minutes. Samples were incubated with the primary antibody at room temperature for 1 hour, incubated with a biotinylated secondary antibody for 30 minutes followed by HRP-Streptavidin for 30 minutes. Developed with DAB chromogen substrate for 5-10 minutes. Counter stained with hematoxylin. Magnification: 10X.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labelling CD3 with ab5690 at 2µl/ml. Slides were steamed in IHC epitope retrieval solution for 35 minutes and then cooled for 20 minutes. Samples were incubated with the primary antibody at room temperature for 1 hour, incubated with a biotinylated secondary antibody for 30 minutes followed by HRP-Streptavidin for 30 minutes. Developed with DAB chromogen substrate for 5-10 minutes. Counter stained with hematoxylin. Magnification: 40X.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse spleen tissue labelling CD3 with ab5690 at 2µl/ml. Slides were steamed in IHC epitope retrieval solution for 35 minutes and then cooled for 20 minutes. Samples were incubated with the primary antibody at room temperature for 1 hour, incubated with a biotinylated secondary antibody for 30 minutes followed by HRP-Streptavidin for 30 minutes. Developed with DAB chromogen substrate for 5-10 minutes. Counter stained with hematoxylin. Magnification: 10X.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse spleen tissue labelling CD3 with ab5690 at 2µl/ml. Slides were steamed in IHC epitope retrieval solution for 35 minutes and then cooled for 20 minutes. Samples were incubated with the primary antibody at room temperature for 1 hour, incubated with a biotinylated secondary antibody for 30 minutes followed by HRP-Streptavidin for 30 minutes. Developed with DAB chromogen substrate for 5-10 minutes. Counter stained with hematoxylin. Magnification: 40X.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human skeletal muscle tissue showing no expression of CD3 when labelled with ab5690 at 2µl/ml. Slides were steamed in IHC epitope retrieval solution for 35 minutes and then cooled for 20 minutes. Samples were incubated with the primary antibody at room temperature for 1 hour, incubated with a biotinylated secondary antibody for 30 minutes followed by HRP-Streptavidin for 30 minutes. Developed with DAB chromogen substrate for 5-10 minutes. Counter stained with hematoxylin. Magnification: 40X.

  • ab5690 staining CD3 in the lymphatic nodule of rat spleen by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections). Tissue was fixed with formaldehyde and a heat mediated antigen retrieval step was performed using citrate buffer pH 6.0. Samples were then blocked with 1% BSA for 25 minutes at 20°C followed by incubation with the primary antibody at a 1/100 dilution for 1 hour at 20°C. A biotin-conjugated goat anti-rat polyclonal was used undiluted as the secondary antibody. CD3 positive cells are mainly distributed in PALS (T-cell region), while sporadic CD3 positive cells are identified in lymphoid follicle and germinal center (B-cell region).

    See Abreview

  • ab5690 staining CD3 in human lymph node tissue by Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded tissue sections). The sections were fixed in paraformaldehyde and subjected to heat-mediated antigen retrieval in citric buffer pH 6.0, prior to blocking with 10% serum for 1 hour at 20°C. The primary antibody was diluted 1/100 and incubated with the sample for 12 hours at 4°C. An HRP-conjugated goat anti-rabbit polyclonal was used as the secondary antibody, diluted 1/200.

    See Abreview

  • ab5690 staining CD3 (green) in mouse lymph node tissue sections by Immunohistochemistry (IHC-Fr - frozen sections). Tissue was fixed with acetone/alcohol and blocked with 1% BSA for 1 hour at 23°C. Samples were incubated with primary antibody (1/250 in 1% BSA) for 12 hours at 4°C. An Alexa Fluor® 568-conjugated goat anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody.

    CD8 (red) was labelled with ab22378 followed by an Alexa Fluor® 647-conjugated goat anti-rabbit IgG.

    CD3+ CD8+ are yellow.

    See Abreview

  • No positive immunostaining for CD3ε, a pan T cell marker (ab5690), was detected in the corneas of scrambled siRNA-treated mice (A) at 5 days p.i. In contrast, positive immunostaining (red) was observed in the peripheral cornea of HIF-1α silenced animals (B). The control sections shown in (C) and (D) were immunostained with species-specific IgG and were positive for SYTOX Green nuclear stain only. Images shown are representative of three independent experiments each with three mice per group. Magnification = 180×; inset = 335×.

参考文献

This product has been referenced in:
  • Danturti S  et al. CD4+ T lymphocytes produce adiponectin in response to transplants. JCI Insight 2:N/A (2017). IHC-P . Read more (PubMed: 28614792) »
  • Connolly NP  et al. Genetically engineered rat gliomas: PDGF-driven tumor initiation and progression in tv-a transgenic rats recreate key features of human brain cancer. PLoS One 12:e0174557 (2017). IHC-Fr ; Rat . Read more (PubMed: 28358926) »

See all 161 Publications for this product

レビューと Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Lung and spleen (multi-block))
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Dako high pH buffer
Permeabilization
No
Specification
Lung and spleen (multi-block)
Blocking step
Vector Casein block as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 24°C
Fixative
Paraformaldehyde
Username

Dr. Henry King

Verified customer

投稿 Aug 07 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Jurkat)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
10 µg
Specification
Jurkat
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Abcam user community

Verified customer

投稿 Jun 13 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rhesus monkey Tissue sections (Brain Tissue)
Specification
Brain Tissue
Username

Eli Shobin

Verified customer

投稿 Dec 07 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Antigen retrieval step
Heat mediated
Sample
Mouse Tissue sections (skin)
Specification
skin
Permeabilization
No
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

投稿 Feb 26 2015

Application
Immunohistochemistry (Frozen sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2.5% · Temperature: 24°C
Sample
Mouse Tissue sections (Spleen)
Specification
Spleen
Permeabilization
Yes - Triton X-100 (0.3%)
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Feb 10 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
DakoCytomation x0909 as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: RT°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH6.0
Sample
Mouse Tissue sections (Spleen)
Specification
Spleen
Permeabilization
No
Fixative
Formaldehyde
Username

Hongwei Shao

Verified customer

投稿 Jan 21 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Normal horse blocking serum as blocking agent for 30 minute(s) · Concentration: 2.5% · Temperature: 25°C
Antigen retrieval step
Other
Sample
Rat Tissue sections (Alveolar bone)
Specification
Alveolar bone
Permeabilization
No
Fixative
Formaldehyde
Username

Ms. Dong-Eun Lee

Verified customer

投稿 Oct 15 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 27°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris HCl ph10
Sample
Mouse Tissue sections (Synovium)
Specification
Synovium
Permeabilization
No
Fixative
zinc buffered formalin
Username

Abcam user community

Verified customer

投稿 Aug 08 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Antigen retrieval step
Heat mediated
Sample
Mouse Tissue sections (Colon)
Specification
Colon
Permeabilization
No
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

投稿 Feb 05 2014

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
Antibody Block as blocking agent for 10 minute(s) · Concentration: 100%
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: CC1 Mild
Sample
Human Tissue sections (Colon)
Specification
Colon
Permeabilization
No
Fixative
10% NBF
Username

Abcam user community

Verified customer

投稿 Dec 16 2013

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

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