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, ICC/IF, Dot blot, WB, IP
- Reacts with: Human
Product nameAnti-CD3 zeta (phospho Y83) antibody [EP776(2)Y]
See all CD3 zeta primary antibodies
DescriptionRabbit monoclonal [EP776(2)Y] to CD3 zeta (phospho Y83)
SpecificityThis antibody detects CD3 zeta phosphorylated on tyrosine 83.
Tested applicationsSuitable for: Flow Cyt, ICC/IF, Dot blot, WB, IPmore details
Unsuitable for: IHC-P
Species reactivityReacts with: Human
Synthetic peptide within Human CD3 zeta aa 50-150 (phospho Y83). The exact sequence is proprietary.
Database link: P20963
- WB: Jurkat whole cell lysate (ab7899).
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.40
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol, 9.85% Tris glycine, 50% PBS
Concentration information loading...
PurityProtein A purified
- Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (Alexa Fluor® 488) (ab237451)
- Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (Alexa Fluor® 647) (ab237452)
- Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] (FITC) (ab237453)
- Anti-CD3 zeta (phospho Y83) antibody [EP776(2)Y] - BSA and Azide free (ab238955)
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.
|Flow Cyt||Use at an assay dependent concentration.|
|ICC/IF||1/100 - 1/250.|
|Dot blot||Use at an assay dependent concentration.|
|WB||1/5000 - 1/10000. Detects a band of approximately 18-22 kDa (predicted molecular weight: 18 kDa).|
FunctionProbable role in assembly and expression of the TCR complex as well as signal transduction upon antigen triggering.
Involvement in diseaseDefects 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 similaritiesBelongs to the CD3Z/FCER1G family.
Contains 3 ITAM domains.
DomainThe ITAM domains mediate interaction with SHB.
modificationsPhosphorylated on Tyr residues after T-cell receptor triggering.
- Information by UniProt
- 4930549J05Rik antibody
- A430104F18Rik antibody
- AW552088 antibody
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.
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.
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 minutes
Blocking buffer 5% NFDM/TBST
Diluting buffer 5% NFDM/TBST
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.
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 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.
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.
ab68236 has been referenced in 11 publications.
- Ramello MC et al. An immunoproteomic approach to characterize the CAR interactome and signalosome. Sci Signal 12:N/A (2019). PubMed: 30755478
- Quintarelli C et al. Choice of costimulatory domains and of cytokines determines CAR T-cell activity in neuroblastoma. Oncoimmunology 7:e1433518 (2018). PubMed: 29872565
- Bustos-Morán E et al. Microtubule-associated protein-4 controls nanovesicle dynamics and T cell activation. J Cell Sci 130:1217-1223 (2017). PubMed: 28209780
- 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
- Brownlie RJ et al. Resistance to TGFß suppression and improved anti-tumor responses in CD8+ T cells lacking PTPN22. Nat Commun 8:1343 (2017). Flow Cyt ; Mouse . PubMed: 29116089