Recombinant 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
-
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.
-
Positive control
- Jurkat whole cell lysate (ab7899)
-
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.
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
-
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
-
Alternative Versions
-
Isotype control
-
Positive Controls
-
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.
|
|
ICC/IF | (1) |
1/250 - 1/500.
|
WB |
1/500 - 1/2000. Detects a band of approximately 21 kDa (predicted molecular weight: 19 kDa).
|
|
IP |
1/20 - 1/50.
|
Notes |
---|
Flow Cyt (Intra)
1/120. |
ICC/IF
1/250 - 1/500. |
WB
1/500 - 1/2000. Detects a band of approximately 21 kDa (predicted molecular weight: 19 kDa). |
IP
1/20 - 1/50. |
Target
-
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
-
Database links
- Entrez Gene: 919 Human
- Omim: 186780 Human
- SwissProt: P20963 Human
- Unigene: 156445 Human
-
Alternative names
- 4930549J05Rik antibody
- A430104F18Rik antibody
- AW552088 antibody
see all
Images
-
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 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.
-
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. -
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).
Protocols
Datasheets and documents
-
SDS download
-
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