概述

  • 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
  • 经测试应用
    适用于: Flow Cyt, IHC-Fr, IHC-P, WB, IHC-FoFrmore details
  • 种属反应性
    与反应: Mouse, Rat, Human, Pig
    预测可用于: Sheep, Rabbit, Horse, Chicken, Cow, Cat, Dog, Baboon, Woodchuck
  • 免疫原

    Synthetic peptide corresponding to CD3.

  • 表位
    aa 156-168 of epsilon chain of Human CD3 protein (intracytoplasmic).
  • 阳性对照
    • WB: Recombinant Human CD3 epsilon protein (ab114153), Jurkat whole cell lysate. Human, mouse and rat thymus tissue lysate. IHC-P: Pig and rat spleen tissue. Human tonsil tissue. Mouse epidydimal fat pad tissue. Rat infarcted heart tissue. Flow Cytometry: Human peripheral blood lymphocytes. Jurkat cells. IHC-Fr: Mouse brain tissue.
  • 常规说明

    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.

性能

应用

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.

应用 Ab评论 说明
Flow Cyt 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

We recommend using Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody.

IHC-Fr Use at an assay dependent concentration. PubMed: 18658050
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 1/200. Predicted molecular weight: 23 kDa.
IHC-FoFr Use at an assay dependent concentration.

靶标

  • 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
  • 数据库链接
  • 别名
    • 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

图片

  • 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

  • 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.

  • 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 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 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 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.

  • 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 ( Goat anti-rabbit biotin) , streptavidin-Alexa 594 was applied.

    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

  • 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.

    See Abreview

  • 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.

文献

This product has been referenced in:
  • Luo N  et al. DNA methyltransferase inhibition upregulates MHC-I to potentiate cytotoxic T lymphocyte responses in breast cancer. Nat Commun 9:248 (2018). Read more (PubMed: 29339738) »
  • Höing B  et al. Stromal versus tumoral inflammation differentially contribute to metastasis and poor survival in laryngeal squamous cell carcinoma. Oncotarget 9:8415-8426 (2018). IHC-P . Read more (PubMed: 29492204) »
See all 160 Publications for this product

客户评价及客户问答

1-10 of 71 Abreviews or 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

Dr. Ling Guo

Verified customer

提交于 Feb 10 2015

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Spleen)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer pH 6
Permeabilization
No
Specification
Spleen
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

提交于 Nov 27 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Frozen sections)
Sample
Human Tissue sections (Human skin)
Permeabilization
Yes - 0.5% Triton TBS Ca/Azide
Specification
Human skin
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative
Formaldehyde

Mr. Tyler Crowe

Verified customer

提交于 Aug 20 2018

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 (Lymph node)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH9
Permeabilization
Yes - Tween-20
Specification
Lymph node
Blocking step
RTU Power Vision blocking from Leica as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: RT°C
Fixative
Formaldehyde

Abcam user community

Verified customer

提交于 Jun 25 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Lymph node)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH 9.0 EDTA
Permeabilization
No
Specification
Lymph node
Blocking step
PB ab64226 as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Formaldehyde

Abcam user community

Verified customer

提交于 Feb 06 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Mouse Epididymal Fat Pad)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate
Permeabilization
Yes - Triton X-100
Specification
Mouse Epididymal Fat Pad
Blocking step
Serum as blocking agent for 4 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Formaldehyde

Ms. Ying Li

Verified customer

提交于 Nov 23 2017

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

Abcam user community

Verified customer

提交于 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

Abcam user community

Verified customer

提交于 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

Dr. Li Li

Verified customer

提交于 May 16 2017

Answer

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 concentration is usually determined by protein assay, and serum / ascites / tissue culture supernatant will contain a lot of other proteins, which means the antibody quantification would not be accurate.


I can confirm that for tissue culture supernatant, concentration of antibody is known to vary between 1 - 3 mg/ml


I am sorry we are not able to provide an exact concentration on this occasion, but hope this information will be helpful to you.



Read More

1-10 of 71 Abreviews or Q&A

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

注册