- Clone
- 1503310 (See other available formats)
- Regulatory Status
- RUO
- Other Names
- Phospho Zeta-Chain (TCR) Associated Protein Kinase 70 kD, Phospho SRK, Phospho 70 KD Zeta-Chain Associated Protein, Phospho Tyrosine-protein kinase ZAP-70
- Isotype
- Mouse IgG1, κ
- Ave. Rating
- Submit a Review
- Product Citations
- publications
![1-1503310_PURE_ZAP70_Phospho_Antibody_1_WB_030816 1-1503310_PURE_ZAP70_Phospho_Antibody_1_WB_030816](https://d1spbj2x7qk4bg.cloudfront.net/Admin/Public/GetImage.ashx?Image=/Files/Images/media_assets/products/product_images/1503310_PURE_ZAP70_Phospho_Antibody_1_WB_030816.jpg&Width=240&Height=300&altFmImage_path=&Compression=90&Crop=5)
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Jurkat cells were treated with 2 mM H2O2 for 15 minutes. Protein (15 µg) extracted from Jurkat cells were resolved by electrophoresis, transferred to nitrocellulose, and probed with purified monoclonal anti-ZAP70 Phospho (Tyr319) (clone 1503310) antibody. Proteins were visualized using a HRP Goat anti-mouse IgG (minimal x-reactivity) antibody and chemiluminescence detection. -
HO treated Jurkat cells were fixed with 4% paraformaldehyde (PFA), permeabilized with 0.5% Triton X‐100, and blocked with 5% FBS. Then the cells were stained with 2 µg/ml anti‐ZAP70 Phospho (Tyr319) (clone 1503310) antibody followed by 2 µg/ml Alexa Fluor® 594 Goat anti‐mouse IgG (minimal x‐reactivity) Antibody. Nuclei were counterstained with DAPI (blue). The image was captured by a 60X objective. -
Human peripheral blood lymphocytes were treated with 0.03% H2O2 (left) or without (right) for 5 mins, followed by fixation with Fixation Buffer (Cat No. 420801), and permeabilization with True-Phos™ Perm Buffer (Cat No. 425401). Cells were stained with CD20 APC and purified ZAP70 Phospho (Tyr319)/Syk Phospho (Tyr352) (clone 1503310), followed by mouse IgG1 PE. -
C57BL/6 splenocytes were treated with 0.03% H2O2 (left) or without (right) for 5 mins, followed by fixation with Fixation Buffer (Cat. No. 420801), and permeabilization with True-Phos™ Perm Buffer (Cat. No. 425401). Cells were stained with CD45R/B220 Brilliant Violet 421™ and purified ZAP70 Phospho (Tyr319)/Syk Phospho (Tyr352) (clone 1503310), followed by anti-mouse IgG1 PE.
Cat # | Size | Price | Quantity Check Availability | Save | ||
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683702 | 100 µg | 265 CHF |
ZAP70 was identified in TCR-stimulated Jurkat cells. It is an inactive cytosolic tyrosine kinase that is recruited to a transmembrane receptor lacking intrinsic catalytic activity. ZAP70 and the related spleen tyrosine kinase (Syk) play a critical role in T-cell development and activation. This enzyme, which is phosphorylated on tyrosine residues upon T-cell antigen receptor (TCR) stimulation, functions in the initial step of TCR-mediated signal transduction in combination with the Src family kinases, Lck and Fyn.
ZAP70 activation can be regulated by binding to phosphorylated ITAMs of the TCR and by phosphorylation of multiple tyrosine residues on ZAP70. Phosphorylation of Tyr-315 and Tyr-319 are essential for ZAP70 positive regulation of T-lymphocyte activation whereas Tyr-292 has a negative regulatory role.
Phosphorylated Y319 is a positive regulator of ZAP70, which triggers the binding of Lck and activation of NFAT and IL-2 induction. The binding of Lck to phosphorylated Y319 promotes Lck mediated phosphorylation of Y493, and facilitates activation of downstream signaling. The importance of Y319 phosphorylation in positive regulation of ZAP70 was confirmed in a mouse model with Y319F ZAP70 mutant expression, which resulted in severe defects in calcium mobilization and thymocyte selection.
Product DetailsProduct Details
- Verified Reactivity
- Human
- Antibody Type
- Monoclonal
- Host Species
- Mouse
- Immunogen
- Modified peptide.
- Formulation
- Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide.
- Preparation
- The antibody was purified by affinity chromatography.
- Concentration
- 0.5 mg/ml
- Storage & Handling
- The antibody solution should be stored undiluted between 2°C and 8°C.
- Application
-
WB - Quality tested
ICC, ICFC - Verified - Recommended Usage
-
Each lot of this antibody is quality control tested by Western blotting. For Western blotting, the suggested use of this reagent is 0.5 - 2.5 µg/ml. For immunofluorescence microscopy, a concentration range of 0.5 - 2.5 μg/ml is recommended. For intracellular flow cytometry using our True-Phos™ Perm Buffer, the suggested use of this reagent is 5 µl per million cells or 5 µl per 100 µl of whole blood. It is recommended that the reagent be titrated for optimal performance for each application.
- Application Notes
-
This antibody can also weakly react with Syk Phospho (Tyr352) on WB.
- Product Citations
-
- RRID
-
AB_2572026 (BioLegend Cat. No. 683702)
Antigen Details
- Structure
- 619 amino acids with a predicted molecular weight of 70 kD.
- Distribution
-
Cytoplasm and cell membrane.
- Function
- Required for the assembly of a Zap-70-containing signaling complex. Activation of the PLC-g1-dependent and Ras-dependent signaling cascades. Association with PLC-g1.
- Interaction
- NFAM1, SLA and SLA2, CBLB, DEF6, FCRL3, VAV1, and CD247/CD3Z.
- Cell Type
- B cells
- Biology Area
- Cell Biology, Immunology, Signal Transduction
- Molecular Family
- Phospho-Proteins, Protein Kinases/Phosphatase, TCRs
- Antigen References
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1. Chan AC, et al. 1991. Proc. Natl. Acad. Sci. 88:9166.
2. Arpaia E, et al. 1994. Cell 76:947.
3. Chan AC, et al. 1994. Science 264:1599.
4. Negishi I, et al. 1995. Nature 376:435.
5. Wang H, et al. 2010. Cold Spring Harb. Perspect. Biol. 2:a002279.
6. Klammt C, et al. 2015. Nat. Immunol. 16:961.
7. Weiss A. 1993. Cell 73:209.
8. Gong Q, et al. 2001. J. Exp. Med. 194:507. - Gene ID
- 7535 View all products for this Gene ID
- UniProt
- View information about ZAP-70 Phospho Tyr319 Syk Phospho Tyr352 on UniProt.org
Related Pages & Pathways
Pages
Other Formats
View All ZAP70 Phospho (Tyr319)/Syk Phospho (Tyr352) Reagents Request Custom ConjugationDescription | Clone | Applications |
---|---|---|
Purified anti-ZAP70 Phospho (Tyr319)/Syk Phospho (Tyr352) | 1503310 | WB,ICC,ICFC |
PE/Cyanine7 anti-ZAP70 Phospho (Tyr319)/Syk Phospho (Tyr352) | 1503310 | ICFC |
Alexa Fluor® 647 anti-ZAP70 Phospho (Tyr319)/Syk Phospho (Tyr352) | 1503310 | ICFC |
Alexa Fluor® 488 anti-ZAP70 Phospho (Tyr319)/Syk Phospho (Tyr352) | 1503310 | ICFC |
PerCP/Cyanine5.5 anti-ZAP70 Phospho (Tyr319)/Syk Phospho (Tyr352) | 1503310 | ICFC |
PE anti-ZAP70 Phospho (Tyr319)/Syk Phospho (Tyr352) | 1503310 | ICFC |
Alexa Fluor® 594 anti-ZAP70 Phospho (Tyr319)/Syk Phospho (Tyr352) | 1503310 | ICC |
Compare Data Across All Formats
This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
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Purified anti-ZAP70 Phospho (Tyr319)/Syk Phospho (Tyr352)
Jurkat cells were treated with 2 mM H2O2 HO treated Jurkat cells were fixed w... Human peripheral blood lymphocytes were treated with 0.03% ... C57BL/6 splenocytes were treated with 0.03% H2O2 (left) or w... -
PE/Cyanine7 anti-ZAP70 Phospho (Tyr319)/Syk Phospho (Tyr352)
Human peripheral blood lymphocytes were treated with (left),... -
Alexa Fluor® 647 anti-ZAP70 Phospho (Tyr319)/Syk Phospho (Tyr352)
Human peripheral blood lymphocytes were treated with (left),... -
Alexa Fluor® 488 anti-ZAP70 Phospho (Tyr319)/Syk Phospho (Tyr352)
Human peripheral blood lymphocytes were treated with (left),... -
PerCP/Cyanine5.5 anti-ZAP70 Phospho (Tyr319)/Syk Phospho (Tyr352)
Human peripheral blood lymphocytes were treated with (left),... -
PE anti-ZAP70 Phospho (Tyr319)/Syk Phospho (Tyr352)
Human PBMCs were treated with (left), or without (right) H Mouse spleen cells were treated with 0.03% H2O2 for five min... -
Alexa Fluor® 594 anti-ZAP70 Phospho (Tyr319)/Syk Phospho (Tyr352)
ICC staining of Alexa Fluor® 594 anti-ZAP70 Phospho (Tyr319)...
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