Product nameAnti-ZAP70 (phospho Y292) antibody [EPR1073]
See all ZAP70 primary antibodies
DescriptionRabbit monoclonal [EPR1073] to ZAP70 (phospho Y292)
Specificityab76501 only detects ZAP70 when phosphorylated at Tyrosine 292.
Tested applicationsSuitable for: WB, Flow Cyt, ICC/IF, Dot blotmore details
Unsuitable for: IHC-P or IP
Species reactivityReacts with: Human
Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human ZAP70 (phospho Y292). The exact sequence is proprietary.
- WB: Jurkat whole cell lysate (ab7899) treated with pervanadate. ICC/IF: Untreated and pervanadate treated Jurkat cells. Flow Cyt: Untreated and pervanadate treated Jurkat cells.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.
This product is a recombinant rabbit monoclonal antibody.
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: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol, 9.85% Tris glycine, 50% Tissue culture supernatant
PurityTissue culture supernatant
Our Abpromise guarantee covers the use of ab76501 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|WB||1/5000 - 1/10000. Predicted molecular weight: 70 kDa.|
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
|ICC/IF||1/250 - 1/500.|
FunctionPlays a role in T-cell development and lymphocyte activation. Essential for TCR-mediated IL-2 production. Isoform 1 induces TCR-mediated signal transduction, isoform 2 does not.
Tissue specificityExpressed in T- and natural killer cells.
Involvement in diseaseDefects in ZAP70 are the cause of selective T-cell defect (STD) [MIM:176947]. STD is an autosomal recessive form of severe combined immunodeficiency characterized by a selective absence of CD8-type T-cells.
Sequence similaritiesBelongs to the protein kinase superfamily. Tyr protein kinase family. SYK/ZAP-70 subfamily.
Contains 1 protein kinase domain.
Contains 2 SH2 domains.
DomainThe SH2 domains bind to the phosphorylated tyrosine-based activation motif (TAM) of CD3Z and the non-canonical phosphorylated tyrosine-based activation motif (TAM) of RHOH.
modificationsPhosphorylated on tyrosine residues upon T-cell antigen receptor (TCR) stimulation. Tyr-319 phosphorylation is essential for full activity.
Cellular localizationCytoplasm. Cell membrane. After antigen stimulation, isoform 1 concentrates at the immunological synapse and isoform 2 remains cytoplasmic. Co-localizes together with RHOH in the immunological synapse. RHOH is required for its proper localization to the cell membrane and cytoskeleton fractions in the thymocytes.
- Information by UniProt
- 70 kDa zeta associated protein antibody
- 70 kDa zeta-associated protein antibody
- EC 220.127.116.11 antibody
All lanes : Anti-ZAP70 (phospho Y292) antibody [EPR1073] (ab76501) at 1/5000 dilution
Lane 1 : Untreated Jurkat (human acute T cell leukemia) whole cell lysates
Lane 2 : Jurkat (human acute T cell leukemia) treated with pervanadate whole cell lysates
Lysates/proteins at 10 µg per lane.
All lanes : Peroxidase conjugated goat anti-rabbit IgG, (H+L) at 1/1000 dilution
Predicted band size: 70 kDa
Observed band size: 70 kDa
Exposure time: 3 minutes
Blocking and dilution buffer: 5% NFDM/TBST.
Dot blot analysis of varying amounts of ZAP70 (pY292) phospho peptide (Lane 1) and ZAP70 non-phospho peptide (Lane 2) using ab76501 at a dilution of 1/1000. A peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/2500).
Exposure time: 3 minutes.
Immunocytochemistry/Immunofluorescence analysis of untreated and pervanadate treated Jurkat (human acute T cell leukemia) cells labelling ZAP70 (phospho Y292) with ab76501 at a dilution of 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Treated samples were starved overnight followed by incubation with 1mM pervanadate for 30 minutes. 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. Negative control using secondary antibody only.
Image shows increased cytoplasmic staining after pervanadate treatment.
Flow cytometric analysis of permeabilized Jurkat cells, untreated (green), pervanadate treated (red) stained with ab76501, at 1/20 dilution (blue) and pervanadate-treated Jurkat cells using the same antibody preincubated with phospho- ZAP70 (Tyr 292) peptide (blue) or non-phospho- ZAP70 (Tyr 292)).
ab76501 has not yet been referenced specifically in any publications.