Recombinant Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)
![Flow Cytometry (Intracellular) - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)](https://www.abcam.com/ps/products/68/ab68236/Images/ab68236-273109-ab68236FC1.jpg "Flow Cytometry (Intracellular) - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP776(2)Y] to CD3 zeta (phospho Y83)
- Suitable for: Flow Cyt (Intra), ICC/IF, Dot blot, WB, IP
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y]
See all CD3 zeta primary antibodies -
Description
Rabbit monoclonal [EP776(2)Y] to CD3 zeta (phospho Y83) -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, Dot blot, WB, IPmore details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide within Human CD3 zeta aa 50-150 (phospho Y83). The exact sequence is proprietary.
Database link: P20963 -
Positive control
- WB: Jurkat whole cell lysate (ab7899). IP: Jurkat. ICC: Jurkat cells. Flow Cyt (intra): Jurkat cells.
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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. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% PBS -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP776(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 ab68236 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) |
Use at an assay dependent concentration.
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| ICC/IF |
1/100 - 1/250.
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| Dot blot |
Use at an assay dependent concentration.
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| WB |
1/5000 - 1/10000. Detects a band of approximately 18-22 kDa (predicted molecular weight: 18 kDa).
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| IP |
1/50.
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| Notes |
|---|
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Flow Cyt (Intra)
Use at an assay dependent concentration. |
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ICC/IF
1/100 - 1/250. |
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Dot blot
Use at an assay dependent concentration. |
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WB
1/5000 - 1/10000. Detects a band of approximately 18-22 kDa (predicted molecular weight: 18 kDa). |
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IP
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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Flow Cytometry (Intracellular) - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)
Flow Cytometry analysis of Jurkat (human acute T cell leukemia) treated (Red)/untreated (Green) with 1mM pervanadate for 4 hours with purified ab68236 at 1/250 dilution. The secondary antibody was Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution. A Rabbit monoclonal IgG (Black) was used as the isotype control and cells without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.
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![Western blot - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)](https://www.abcam.com/ps/products/68/ab68236/Images/ab68236-258230-ab68236WB.jpg)
Western blot - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)All lanes : Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236) at 1/2000 dilution
Lane 1 : Untreated Jurkat cells whole cell lysates
Lane 2 : Jurkat cells were treated with 50mM Pervanadate for 5 minutes whole cell lysates
Lane 3 : Jurkat cells were treated with 50mM Pervanadate for 5 minutes whole cell lysates. Then the membrane was incubated with Alkaline phosphatase.
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 18 kDa
Observed band size: 18 kDa
Exposure time: 3 minutesBlocking buffer 5% NFDM/TBST
Diluting buffer 5% NFDM/TBST
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![Western blot - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)](https://www.abcam.com/ps/products/68/ab68236/Images/ab68236.jpg)
Western blot - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)All lanes : Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236) at 1/10000 dilution
Lane 1 : Jurkat cell lysate, untreated.
Lane 2 : Jurkat cell lysate, treated with pervanadate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution
Predicted band size: 18 kDa
Observed band size: 18-22 kDa why is the actual band size different from the predicted? -
![Immunocytochemistry/ Immunofluorescence - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)](https://www.abcam.com/ps/products/68/ab68236/Images/ab68236-255370-ab68236iccif.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)Immunocytochemistry/Immunofluorescence analysis of Jurkat cells (untreated, Per treated and Per+LP treated) labelling CD3 zeta (phospho Y83) with ab68236 (left) and CD3 zeta with ab40804 (right) both at a dilution of 1/200. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
The image shows increased cytoplamic staining after Pervanadate (1 mM, 30 min) treatment on Jurkat cells. The LP treatment decreased the cytoplasmic staining caused by Pervanadate.
ab40804 was used as a Pan control for ab68236. The results showed cytoplamic staining on untreated, pervanadate (1 mM, 30 min) treated and Per+LP treated Jurkat cells.
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![Immunoprecipitation - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)](https://www.abcam.com/ps/products/68/ab68236/Images/ab68236-7-anti-cd3-zeta-phospho-y83-antibody-ep776-2-y-immunoprecipitation-jurkat-human.jpg)
Immunoprecipitation - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)CD3 zeta was immunoprecipitated from 0.35 mg Jurkat (Human T cell leukemia T lymphocyte) treated with pervandate (50mM 5min) whole cell lysate 10 µg with ab68236 at 1/30 dilution (2µg) . VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.
Lane 1: Jurkat (Human T cell leukemia T lymphocyte) treated with pervandate (50mM 5min) whole cell lysate 10 µg
Lane 2: ab68236 IP in Jurkat treated with pervandate (50mM 5min) whole cell lysate
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab68236 in Jurkat treated with pervandate (50mM 5min) whole cell lysate
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
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![Dot Blot - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)](https://www.abcam.com/ps/products/68/ab68236/Images/ab68236-253367-ab68236dot.jpg)
Dot Blot - Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)Dot blot analysis of CD3 zeta (pY83) phospho peptide (lane 1) and CD3 zeta non-phospho peptide (lane 2) labelling CD3 zeta (phospho Y83) with ab68236 at a dilution of 1/1000. A peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/2500).
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure time: 3 minutes.
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![Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)](https://www.abcam.com/ps/products/68/ab68236/Images/ab68236-6-benefits-of-recombinant-antibodies.png)
Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (ab68236)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (14)
ab68236 has been referenced in 14 publications.
- Eleftheriadis T et al. A Role for Human Renal Tubular Epithelial Cells in Direct Allo-Recognition by CD4+ T-Cells and the Effect of Ischemia-Reperfusion. Int J Mol Sci 22:N/A (2021). PubMed: 33572206
- 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
- Ramello MC et al. An immunoproteomic approach to characterize the CAR interactome and signalosome. Sci Signal 12:N/A (2019). PubMed: 30755478
- Razvag Y et al. T Cell Activation through Isolated Tight Contacts. Cell Rep 29:3506-3521.e6 (2019). PubMed: 31825832
- Quintarelli C et al. Choice of costimulatory domains and of cytokines determines CAR T-cell activity in neuroblastoma. Oncoimmunology 7:e1433518 (2018). PubMed: 29872565