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Alexa Fluor® 647 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
1503310_A647_ZAP70_Tyr319_Syk_Tyr352_Antibody_052219_updated.png
Human peripheral blood lymphocytes were treated with (left), or without (right) H2O2 for five minutes, fixed with Fixation Buffer, permeabilized with True-Phos™ Perm Buffer, then surface stained with CD3 Brilliant Violet 421™ and intracellularly stained with anti-ZAP70 Phospho (Tyr319)/Syk (Tyr352) (clone 1503310) Alexa Fluor® 647.
  • 1503310_A647_ZAP70_Tyr319_Syk_Tyr352_Antibody_052219_updated.png
    Human peripheral blood lymphocytes were treated with (left), or without (right) H2O2 for five minutes, fixed with Fixation Buffer, permeabilized with True-Phos™ Perm Buffer, then surface stained with CD3 Brilliant Violet 421™ and intracellularly stained with anti-ZAP70 Phospho (Tyr319)/Syk (Tyr352) (clone 1503310) Alexa Fluor® 647.
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683705 25 tests 136 CHF
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683706 100 tests 347 CHF
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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 and BSA (origin USA)
Preparation
The antibody was purified by affinity chromatography and conjugated with Alexa Fluor® 647 under optimal conditions.
Concentration
Lot-specific (to obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.)
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application

ICFC - Quality tested
Recommended Usage

Each lot of this antibody is quality control tested by intracellular immunofluorescent staining with flow cytometric analysis. For flow cytometric staining, the suggested use of this reagent is 5 µl per million cells in 100 µl staining volume or 5 µl per 100 µl of whole blood.

* Alexa Fluor® 647 has a maximum emission of 668 nm when it is excited at 633 nm / 635 nm.


Alexa Fluor® and Pacific Blue™ are trademarks of Life Technologies Corporation.

View full statement regarding label licenses
Excitation Laser
Red Laser (633 nm)
Application Notes

This antibody can also weakly react with Syk Phospho (Tyr352) on WB.

Product Citations
  1. Yin J, et al. 2023. Front Pharmacol. 1191692:14. PubMed
RRID
AB_2687046 (BioLegend Cat. No. 683705)
AB_2687046 (BioLegend Cat. No. 683706)

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: 0    Revision Date: 05.03.2017

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.
If you need assistance with selecting the best format contact our expert technical support team.

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