Recombinant Anti-ZAP70 (phospho Y292) antibody [EPR1073] (ab76501)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR1073] to ZAP70 (phospho Y292)
- Suitable for: Flow Cyt (Intra), WB, ICC/IF, Dot blot
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-ZAP70 (phospho Y292) antibody [EPR1073]
See all ZAP70 primary antibodies -
Description
Rabbit monoclonal [EPR1073] to ZAP70 (phospho Y292) -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, ICC/IF, Dot blotmore details
Unsuitable for: IHC-P or IP -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Jurkat whole cell lysate (ab7899) treated with pervanadate. ICC/IF: Untreated and pervanadate treated Jurkat cells. Flow Cyt (intra): Untreated and pervanadate treated 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.
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: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR1073 -
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 ab76501 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 |
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Flow Cyt (Intra) |
1/20.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB | (1) |
1/5000 - 1/10000. Predicted molecular weight: 70 kDa.
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ICC/IF |
1/250 - 1/500.
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Dot blot |
1/1000.
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Notes |
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Flow Cyt (Intra)
1/20. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/5000 - 1/10000. Predicted molecular weight: 70 kDa. |
ICC/IF
1/250 - 1/500. |
Dot blot
1/1000. |
Target
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Function
Plays 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 specificity
Expressed in T- and natural killer cells. -
Involvement in disease
Defects 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 similarities
Belongs to the protein kinase superfamily. Tyr protein kinase family. SYK/ZAP-70 subfamily.
Contains 1 protein kinase domain.
Contains 2 SH2 domains. -
Domain
The 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. -
Post-translational
modificationsPhosphorylated on tyrosine residues upon T-cell antigen receptor (TCR) stimulation. Tyr-319 phosphorylation is essential for full activity. -
Cellular localization
Cytoplasm. 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
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Database links
- Entrez Gene: 7535 Human
- Omim: 176947 Human
- SwissProt: P43403 Human
- Unigene: 234569 Human
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Alternative names
- 70 kDa zeta associated protein antibody
- 70 kDa zeta-associated protein antibody
- EC 2.7.10.2 antibody
see all
Images
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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.
Secondary
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 minutesBlocking and dilution buffer: 5% NFDM/TBST.
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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.
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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.
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Intracellular 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)).
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (0)
ab76501 has not yet been referenced specifically in any publications.