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Purified anti-ZAP70 Phospho (Tyr319)/Syk Phospho (Tyr352) Antibody

Pricing & Availability
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
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Product Citations
publications
1-1503310_PURE_ZAP70_Phospho_Antibody_1_WB_030816
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.
  • 1-1503310_PURE_ZAP70_Phospho_Antibody_1_WB_030816
    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.
  • 2-1503310_PURE_ZAP70_Phospho_Antibody_IHC_010518
    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.
  • 3-1503310_Pure_ZAP70_Tyr319_Syk_Tyr352_Antibody_3_043018
    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.
  • 4-1503310_Pure_ZAP70_Tyr319_Syk_Tyr352_Antibody_4_123019_updated
    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.
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683702 100 µg 212€
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Description

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 Details
Technical Data Sheet (pdf)

Product 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
  1. Wu Y, et al. 2021. J Clin Invest. 131:. PubMed
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

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
Go To Top Version: 3    Revision Date: 04-30-2018

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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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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