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Purified anti-ZAP70 Phospho (Tyr292) Antibody

Pricing & Availability
Clone
A16038A (See other available formats)
Regulatory Status
RUO
Other Names
p Zeta-Chain (TCR) Associated Protein Kinase 70 kD, p 70 KD Zeta-Chain Associated Protein, p Tyrosine-protein kinase ZAP-70
Isotype
Mouse IgG2b, κ
Ave. Rating
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Product Citations
publications
A_A16038A_PURE_ZAP70_Phospho_Tyr292_Antibody_WB_110316
Total cell lysates (15 µg protein) from non-treated and 2 mM H2O2 treated (15 minute treatment) Jurkat and EL-4 were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to nitrocellulose, and probed with 0.05 µg/mL purified anti-ZAP70 Phospho (Tyr292) (clone A16038A) antibody (upper), 0.5 µg/mL purified anti-ZAP70 (clone A15114B) antibody (middle). Proteins were visualized using a goat anti-mouse-IgG secondary antibody conjugated to HRP and chemiluminescence detection. Direct-Blot™ HRP anti-β-actin (clone 2F1-1) antibody was used as a loading control (lower).
  • A_A16038A_PURE_ZAP70_Phospho_Tyr292_Antibody_WB_110316
    Total cell lysates (15 µg protein) from non-treated and 2 mM H2O2 treated (15 minute treatment) Jurkat and EL-4 were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to nitrocellulose, and probed with 0.05 µg/mL purified anti-ZAP70 Phospho (Tyr292) (clone A16038A) antibody (upper), 0.5 µg/mL purified anti-ZAP70 (clone A15114B) antibody (middle). Proteins were visualized using a goat anti-mouse-IgG secondary antibody conjugated to HRP and chemiluminescence detection. Direct-Blot™ HRP anti-β-actin (clone 2F1-1) antibody was used as a loading control (lower).
  • A16038B_PURE_ZAP70_Phospho_Antibody_2_052819_updated.png
    Human peripheral blood lymphocytes were treated with (left), or without (right), Hydrogen Peroxide for 5 minutes, fixed with Fixation Buffer (Cat. No. 420801), permeabilized with True-Phos™ Perm Buffer (Cat No. 425401), then surface stained with CD3 (clone OKT3) APC and intracellularly stained with purified anti-ZAP70 Phospho (Tyr292) (clone A16038A). Followed by anti-mouse IgG2b PE.
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691902 100 µg £197
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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.

Mutation of Y292 to phenylalanine enhanced NFAT induction in response to BCR stimulation, indicating phosphorylation of Y292 may promote association of ZAP-70 with a negative regulatory protein. ZAP-70 has been reported to bind Cbl following TCR stimulation and phosphorylation of Y292 was required for this interaction, as well as for the negative effect of Cbl on signaling.

Product Details
Technical Data Sheet (pdf)

Product Details

Verified Reactivity
Human, Mouse
Antibody Type
Monoclonal
Host Species
Mouse
Immunogen
Human ZAP70 peptide phosphorylated at Y292.
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
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 0.05 - 0.5 µg per ml. Each lot of this antibody is quality control tested by intracellular flow cytometry using our True-Phos™ Perm Buffer in Whole Blood Protocol. For flow cytometric staining, 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.

Product Citations
  1. Voisinne G, et al. 2022. Nat Immunol. 23:1355. PubMed
RRID
AB_2632759 (BioLegend Cat. No. 691902)

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-γ1-dependent and Ras-dependent signaling cascades. Associates with PLC-γ 1.
Interaction
NFAM1, Cbl, SLA and SLA2, CBLB, DEF6, FCRL3, VAV1, CD247/CD3Z.
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. USA 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, et al. 1993. Cell. 73:209.
8. Gong Q, et al. 2001. J. Exp. Med. 194:507.
9. Magnan A, et al. 2001. J. Exp. Med. 194:491.

Gene ID
7535 View all products for this Gene ID
UniProt
View information about ZAP-70 Phospho Tyr292 on UniProt.org
Go To Top Version: 2    Revision Date: 12/07/2016

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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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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