Recombinant Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] (ab68235)
![Western blot - Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] (ab68235)](https://www.abcam.com/ps/products/68/ab68235/Images/ab68235-344774-Anti-cd3-zeta-phosphoY142-antibody-EP265-2Y-westernblot-human.jpg "Western blot - Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] (ab68235)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP265(2)Y] to CD3 zeta (phospho Y142)
- Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y]
See all CD3 zeta primary antibodies -
Description
Rabbit monoclonal [EP265(2)Y] to CD3 zeta (phospho Y142) -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, WB, IPmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- Jurkat whole cell lysate (ab7899)
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP265(2)Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab68235 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| Flow Cyt (Intra) |
1/120.
|
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| ICC/IF |
1/250 - 1/500.
|
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| WB |
1/500 - 1/2000. Detects a band of approximately 21 kDa (predicted molecular weight: 19 kDa).
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| IP |
1/20 - 1/50.
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| Notes |
|---|
|
Flow Cyt (Intra)
1/120. |
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ICC/IF
1/250 - 1/500. |
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WB
1/500 - 1/2000. Detects a band of approximately 21 kDa (predicted molecular weight: 19 kDa). |
|
IP
1/20 - 1/50. |
Target
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Function
Probable role in assembly and expression of the TCR complex as well as signal transduction upon antigen triggering. -
Involvement in disease
Defects in CD247 are the cause of immunodeficiency due to defect in CD3-zeta (CD3ZID) [MIM:610163]. An immunological deficiency characterized by T-cells impaired immune response to alloantigens, tetanus toxoid and mitogens. -
Sequence similarities
Belongs to the CD3Z/FCER1G family.
Contains 3 ITAM domains. -
Domain
The ITAM domains mediate interaction with SHB. -
Post-translational
modificationsPhosphorylated on Tyr residues after T-cell receptor triggering. -
Cellular localization
Membrane. - Information by UniProt
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Database links
- Entrez Gene: 919 Human
- Omim: 186780 Human
- SwissProt: P20963 Human
- Unigene: 156445 Human
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Alternative names
- 4930549J05Rik antibody
- A430104F18Rik antibody
- AW552088 antibody
see all
Images
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Western blot - Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] (ab68235)All lanes : Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] (ab68235) at 1/1000 dilution (Purified)
Lane 1 : Jurkat (Human T cell leukemia T lymphocyte) whole cell lysates
Lane 2 : Jurkat (Human T cell leukemia T lymphocyte) treated with 50mM pervanadate for 5 minutes whole cell lysates
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 19 kDa
Observed band size: 19 kDa -
![Immunocytochemistry/ Immunofluorescence - Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] (ab68235)](https://www.abcam.com/ps/products/68/ab68235/Images/ab68235-3-anticd3zetaphosphoY142antibodyEP2652Y-immunocytochemistryimmunofluorescence-jurkat-human.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] (ab68235)Immunocytochemistry/ Immunofluorescence analysis of Jurkat (Human T cell leukemia T lymphocyte) treated with 1mM pervanadate for 30 minutes cells labeling CD3 zeta with purified ab68235 at 1/500 dilution (0.2 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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![Immunoprecipitation - Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] (ab68235)](https://www.abcam.com/ps/products/68/ab68235/Images/ab68235-5-anticd3zetaphosphoY142antibodyEP2652Y-immunoprecipitation-jurkat-human.jpg)
Immunoprecipitation - Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] (ab68235)ab68235 (purified) at 1/20 dilution (0.5ug) immunoprecipitating CD3 zeta in Jurkat treated with 50nM pervanadate for 5 minutes whole cell lysate whole cell lysate. Jurkat (Human T cell leukemia T lymphocyte) treated with 50nM pervanadate for 5 minutes whole cell lysate 10ug
Lane 2 (+): ab68235 & Jurkat treated with 50nM pervanadate for 5 minutes whole cell lysate whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab68235 in Jurkat treated with 50nM pervanadate for 5 minutes whole cell lysate
For western blotting, VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/2000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST. -
![Flow Cytometry (Intracellular) - Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] (ab68235)](https://www.abcam.com/ps/products/68/ab68235/Images/ab68235-4-anticd3zetaphosphoY142antibodyEP2652Y-flowcytometry-jurkat-human.jpg)
Flow Cytometry (Intracellular) - Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] (ab68235)Intracellular Flow Cytometry analysis of Jurkat (Human T cell leukemia T lymphocyte) treated with 50nM pervanadate for 5 minutes cells labeling CD3 zeta with purified ab68235 at 1/120 dilution (1µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluorr® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue). Untreated Jurkat cells (Green).
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![Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] (ab68235)](https://www.abcam.com/ps/products/68/ab68235/Images/ab68235-7-benefits-of-recombinant-antibodies.png)
Anti-CD3 zeta (phospho Y142) antibody [EP265(2)Y] (ab68235)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (9)
ab68235 has been referenced in 9 publications.
- Lanz AL et al. Allosteric activation of T cell antigen receptor signaling by quaternary structure relaxation. Cell Rep 36:109375 (2021). PubMed: 34260912
- Ma Y et al. Clustering of the ?-Chain Can Initiate T Cell Receptor Signaling. Int J Mol Sci 21:N/A (2020). PubMed: 32429097
- Sun C et al. THEMIS-SHP1 Recruitment by 4-1BB Tunes LCK-Mediated Priming of Chimeric Antigen Receptor-Redirected T Cells. Cancer Cell 37:216-225.e6 (2020). PubMed: 32004441
- Caratelli S et al. In vitro elimination of epidermal growth factor receptor-overexpressing cancer cells by CD32A-chimeric receptor T cells in combination with cetuximab or panitumumab. Int J Cancer 146:236-247 (2020). PubMed: 31479522
- Weber EW et al. Pharmacologic control of CAR-T cell function using dasatinib. Blood Adv 3:711-717 (2019). PubMed: 30814055
- Lee J et al. Induction of Immunosuppressive CD8+CD25+FOXP3+ Regulatory T Cells by Suboptimal Stimulation with Staphylococcal Enterotoxin C1. J Immunol 200:669-680 (2018). WB ; Human . PubMed: 29237775
- Schaer DA et al. The CDK4/6 Inhibitor Abemaciclib Induces a T Cell Inflamed Tumor Microenvironment and Enhances the Efficacy of PD-L1 Checkpoint Blockade. Cell Rep 22:2978-2994 (2018). WB . PubMed: 29539425
- Li L et al. Ionic CD3-Lck interaction regulates the initiation of T-cell receptor signaling. Proc Natl Acad Sci U S A 114:E5891-E5899 (2017). PubMed: 28659468
- Pageon SV et al. Functional role of T-cell receptor nanoclusters in signal initiation and antigen discrimination. Proc Natl Acad Sci U S A 113:E5454-63 (2016). ICC/IF . PubMed: 27573839