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Invitrogen
Phospho-ZAP70/Syk (Tyr319, Tyr352) Monoclonal Antibody (n3kobu5), PE-Cyanine7, eBioscience™
FIGURE: 1 / 2
Phospho-ZAP70/Syk (Tyr319, Tyr352) Antibody (25-9006-42) in Flow
C57BL/6 mouse splenocytes were untreated (orange histogram) or treated for 5 minutes with hydrogen peroxide-activated sodium pervanadate. Cells were then stained intracellularly, using the Intracellular Fixation & Permeabilization Buffer Set (Product # 88-8824-00) and protocol, with CD3 Monoclonal Antibody, eFluor 450 (Product # 48-0032-82) and 0.06 µg of Phospho-ZAP70/Syk (Tyr319, Tyr352) Monoclonal Antibo... View More
Product Details
25-9006-42
Applications
Tested Dilution
Publications
Product Specifications
Species Reactivity
Human,
Mouse
Published species
Not Applicable
Host/Isotype
Mouse
/ IgG2b, kappa
Recommended Isotype Control
Class
Monoclonal
Type
Antibody
Clone
n3kobu5
Conjugate
Excitation/Emission Max
569/780 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_2811791
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-diluted and tested by intracellular staining and flow cytometric analysis of sodium pervanadate-treated mouse splenocytes using the Intracellular Fixation & Permeabilization Buffer Set (a href="https://www.thermofisher.com/order/catalog/product/88-8824-00">Product # 88-8824-00) and protocol. Please refer to "Staining Intracellular Antigens for Flow Cytometry, Protocol A: Two step protocol for intracellular (cytoplasmic) proteins" located at www.thermofisher.com/flowprotocols . This may be used at 5 µL (0.06 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.
Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light.
Fixation: Samples can be stored in IC Fixation Buffer (Product # 00-8222-49) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (Product # 00-5333-57) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically.
Excitation: 488-561 nm; Emission: 775 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser.
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.
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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
Gene Aliases: ADMIO2; IMD48; mrtle; mur; SRK; STCD; STD; TZK; ZAP-70; ZAP70
UniProt ID: (Human) P43403, (Mouse) P43404
Entrez Gene ID: (Human) 7535, (Mouse) 22637
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
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
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