Invitrogen

Phospho-ZAP70/Syk (Tyr319, Tyr352) Monoclonal Antibody (n3kobu5), PE, eBioscience™

1 Published Figure

3 References

View all (97) Zap-70 antibodies

Datasheet

Protocols

Questions & Answers

Datasheet

Protocols

Questions & Answers

Antibody Testing Data (1)
Published Figures (1)

Phospho-ZAP70/Syk (Tyr319, Tyr352) Antibody in Flow Cytometry (Flow)

FIGURE: 1 / 2

Phospho-ZAP70/Syk (Tyr319, Tyr352) Antibody (12-9006-42) in Flow

Intracellular staining of untreated (orange histogram) or 5-minute hydrogen peroxide-activated sodium pervanadate-treated (purple histogram) mouse spleen cells with Anti-Human/Mouse phospho-ZAP-70/SYK (Y319/Y352) PE using the Intracellular Fixation & Permeabilization Buffer Set (Product # 88-8824-00) and protocol. Lymphocytes in the CD3+ (left) or B220+ (right) gates were used for analysis.

Product Image Gallery

Product Details

12-9006-42

Applications

Tested Dilution

Publications

Flow Cytometry (Flow)

5 µL (0.03 µg)/test

View 3 publications 3 publications

Product Specifications

Species Reactivity

Human, Mouse

Published species

Mouse

Host/Isotype

Mouse / IgG2b, kappa

Recommended Isotype Control

Mouse IgG2b kappa Isotype Control (eBMG2b), PE, eBioscience™

Class

Monoclonal

Type

Antibody

Clone

n3kobu5

Conjugate

PE PE PE

Excitation/Emission Max

565/576 nm View spectra

Form

Liquid

Concentration

5 µL/Test

Purification

Affinity chromatography

Storage buffer

PBS, pH 7.2, with 0.2% BSA

Contains

0.09% sodium azide

Storage conditions

4° C, store in dark, DO NOT FREEZE!

Shipping conditions

Ambient (domestic); Wet ice (international)

RRID

AB_2572665

Product Specific Information

Description: This n3kobu5 monoclonal antibody recognizes human and mouse zeta chain-associated protein of 70 kD (also known as ZAP-70) and spleen tyrosine kinase (also known as SYK) when phosphorylated on Y319 and Y352, respectively. ZAP-70 and SYK are members of the SYK protein tyrosine kinase (PTK) family that are phosphorylated and activated by Src family PTKs. ZAP-70/SYK Y319/Y352 are located in the so-called interdomain of ZAP-70/SYK (between the N-terminal dual SH2 domains and the C-terminal kinase domain).

Phosphorylation of ZAP-70 Y319 by Lck is necessary for T cell receptor (TCR)-dependent association of ZAP-70 with Lck and phospholipase C gamma and subsequent activation of calcium-dependent and Ras signaling cascades. SYK Y352 phosphorylation by Fyn/Lyn is critical for propagation of B cell receptor (BCR) signaling and for B cell development.

Specificity of this n3kobu5 clone was determined by ELISA, flow cytometry, and western blotting.

Applications Reported: This n3kobu5 antibody has been reported for use in intracellular staining followed by flow cytometric analysis.

Applications Tested: This n3kobu5 antibody has been pre-titrated and tested by intracellular staining followed by flow cytometric analysis of stimulated normal human peripheral blood cells. This can be used at 5 µL (0.03 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test.

Staining Protocol: All protocols work well for this monoclonal antibody. Use of Protocol A: Two-step protocol: intracellular (cytoplasmic) proteins allows for the greatest flexibility for detection of surface and intracellular (cytoplasmic) proteins. Use of Protocol B: One-step protocol: intracellular (nuclear) proteins is recommended for staining of transcription factors in conjunction with surface and phosphorylated intracellular (cytoplasmic) proteins. Protocol C: Two-step protocol: Fixation/Methanol allows for the greatest discrimination of phospho-specific signaling between unstimulated and stimulated samples, but with limitations on the ability to stain specific surface proteins (refer to "Clone Performance Following Fixation/Permeabilization" located in the Best Protocols Section under the Resources tab online). All Protocols can be found in the Flow Cytometry Protocols: "Staining Intracellular Antigens for Flow Cytometry Protocol" located in the Best Protocols Section under the Resources tab online.

Excitation: 488-561 nm; Emission: 578 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.

Filtration: 0.2 µm post-manufacturing filtered.

Target Information

ZAP-70 is a cytosolic protein tyrosine kinase that is a member of the Syk family of proteins. ZAP-70 is expressed exclusively in T-cells and natural killer cells and is required for T-cell receptor activation. Upon T cell antigen receptor (TCR) engagement, ZAP70 is phosphorylated on tyrosines 292, 315 and 319 in the interdomain B, located between the SH2 and kinase domains. Phosphorylation of both tyrosines 315 (a Vav binding site) and 319 (a Lck binding site) enhances ZAP70 function in mediating lymphocyte signaling, while tyrosine 292 terminates the transient activation of ZAP70 and attentuates lymphocyte signaling.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

How to use the Panel Builder

Watch the video to learn how to use the Invitrogen Flow Cytometry Panel Builder to build your next flow cytometry panel in 5 easy steps.

Product References

Long-Term Programming of CD8 T Cell Immunity by Perinatal Exposure to Glucocorticoids.

Cell

Hong JY,Lim J,Carvalho F,Cho JY,Vaidyanathan B,Yu S,Annicelli C,Ip WKE,Medzhitov R

12-9006 was used in Flow cytometry/Cell sorting to demonstrate that perinatal stress can have long-term consequences on CD8 T cell immunity by altering HPA axis activity.

Thu Mar 05 00:00:00 EST 2020

CD19-CAR-T Cells Bearing a KIR/PD-1-Based Inhibitory CAR Eradicate CD19⁺HLA-C1^- Malignant B Cells While Sparing CD19⁺HLA-C1⁺ Healthy B Cells.

Cancers

Tao L,Farooq MA,Gao Y,Zhang L,Niu C,Ajmal I,Zhou Y,He C,Zhao G,Yao J,Liu M,Jiang W

Published figure using Phospho-ZAP70/Syk (Tyr319, Tyr352) monoclonal antibody (Product # 12-9006-42) in Flow Cytometry

Sun Sep 13 00:00:00 EDT 2020

Neurokinin-1 Receptor Signaling Is Required for Efficient Ca²⁺ Flux in T-Cell-Receptor-Activated T Cells.

Cell reports

Morelli AE,Sumpter TL,Rojas-Canales DM,Bandyopadhyay M,Chen Z,Tkacheva O,Shufesky WJ,Wallace CT,Watkins SC,Berger A,Paige CJ,Falo LD,Larregina AT

12-9006 was used in Flow cytometry/Cell sorting to demonstrate an effect of the NK1R in T cells that is relevant for immunotherapies based on pro-inflammatory neuropeptides and its receptors.

Tue Mar 10 00:00:00 EDT 2020

Bioinformatics

Protein Aliases: 70 kDa zeta-associated protein; 70 kDa zeta-chain associated protein; EC 2.7.10.2; FLJ17670; FLJ17679; kinase ZAP70; Selective T cell defect; Syk-related tyrosine kinase; Tyrosine-protein kinase ZAP-70; Tyrosine-protein kinase ZAP-70;TZK; ZA70; Zeta chain associated protein kinase 70kDa; zeta chain of T cell receptor associated protein kinase 70kDa; zeta-chain (TCR) associated protein kinase 70kDa; zeta-chain associated protein kinase, 70kD

View more View less

Gene Aliases: ADMIO2; IMD48; mrtle; mur; SRK; STCD; STD; TZK; ZAP-70; ZAP70

View more View less

UniProt ID: (Human) P43403, (Mouse) P43404

View more View less

Entrez Gene ID: (Human) 7535, (Mouse) 22637

View more View less

Function(s)

phosphotyrosine binding protein tyrosine kinase activity non-membrane spanning protein tyrosine kinase activity receptor binding protein binding ATP binding nucleotide binding protein kinase activity kinase activity transferase activity transferase activity, transferring phosphorus-containing groups

Process(es)

adaptive immune response macrophage activation involved in immune response neutrophil activation involved in immune response protein phosphorylation inflammatory response immune response transmembrane receptor protein tyrosine kinase signaling pathway peptidyl-tyrosine phosphorylation T cell differentiation positive regulation of cell adhesion mediated by integrin intracellular signal transduction peptidyl-tyrosine autophosphorylation T cell activation B cell activation positive regulation of mast cell degranulation beta selection positive thymic T cell selection negative thymic T cell selection innate immune response positive regulation of B cell differentiation positive regulation of T cell differentiation positive regulation of alpha-beta T cell differentiation positive regulation of alpha-beta T cell proliferation positive regulation of calcium-mediated signaling T cell receptor signaling pathway B cell receptor signaling pathway T cell aggregation T cell migration regulation of platelet aggregation immune system process phosphorylation cell migration cell differentiation thymic T cell selection protein autophosphorylation

Performance Guarantee

If an Invitrogen™ antibody doesn't perform as described on our website or datasheet,we'll replace the product at no cost to you, or provide you with a credit for a future purchase.*

Learn more

Phospho-ZAP70/Syk (Tyr319, Tyr352) Monoclonal Antibody (n3kobu5), PE, eBioscience™

Phospho-ZAP70/Syk (Tyr319, Tyr352) Antibody (12-9006-42) in Flow

Product Details

12-9006-42

Flow Cytometry (Flow)

Species Reactivity

Published species

Host/Isotype

Recommended Isotype Control

Class

Type

Clone

Conjugate

Excitation/Emission Max

Form

Concentration

Purification

Storage buffer

Contains

Storage conditions

Shipping conditions

RRID

Product Specific Information

Target Information

How to use the Panel Builder

Bioinformatics

Performance Guarantee

We're here to help

Your items have has been added!