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Phospho-ZAP70/Syk (Tyr319, Tyr352) Monoclonal Antibody (n3kobu5), APC, eBioscience™
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FIGURE: 1 / 2
Phospho-ZAP70/Syk (Tyr319, Tyr352) Antibody (17-9006-41) in Flow
LEFT: Intracellular staining of whole human blood left unstimulated (orange histogram) or stimulated with Anti-Human CD3 Functional Grade Purified (Product # 16-0038-81) and goat anti-mouse IgG for 5 minutes (purple histogram) with Anti-Human/Mouse phospho-ZAP-70/SYK (Y319/Y352) APC. The 1-step Fix/Lyse Solution (Product # 00-5333-54) and Permeabilization Buffer (Product # 00-8333-56) and protocols were use... View More
Product Details
17-9006-41
Applications
Tested Dilution
Publications
Product Specifications
Published species
Human,
Mouse
Host/Isotype
Mouse
/ IgG2b, kappa
Recommended Isotype Control
Class
Monoclonal
Type
Antibody
Clone
n3kobu5
Conjugate
Excitation/Emission Max
651/660 nm
View spectra
Form
Liquid
Concentration
5 µL/Test
Storage conditions
4°C, store in dark, DO NOT FREEZE!
Shipping conditions
Wet ice
RRID
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 BestProtocols 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 BestProtocols® Section under the Resources tab online.
Excitation: 633-647 nm; Emission: 660 nm; Laser: Red Laser.
Filtration: 0.2 µm post-manufacturing filtered.
Target Information
ZAP70 and SYK are closely related cytoplasmic tyrosine kinases that play central roles in immune receptor signaling. ZAP70 (Zeta-chain-associated protein kinase 70; gene: ZAP70) is primarily expressed in T lymphocytes and natural killer (NK) cells, where it is recruited to the T cell receptor (TCR) complex following antigen recognition. Upon activation, ZAP70 phosphorylates downstream adaptor proteins and initiates signaling cascades that drive T cell activation, proliferation, and differentiation. SYK (Spleen Tyrosine Kinase; gene: SYK) is predominantly expressed in B cells and other hematopoietic cells and functions analogously in B cell receptor (BCR) signaling, as well as in Fc receptor and integrin-mediated pathways. Although ZAP70 and SYK share structural homology and overlapping signaling mechanisms, they exhibit distinct expression patterns and immunological roles. Dysregulation of either kinase has been implicated in autoimmune disease, immunodeficiency, and hematologic malignancies, making them important targets in immunology and therapeutic research.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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