Thermo Fisher Scientific Phospho-ZAP70/Syk (Tyr319, Tyr352) Monoclonal Antibody (n3kobu5), APC, eBioscience

Thermo Fisher Scientific Phospho-ZAP70/Syk (Tyr319, Tyr352) Monoclonal Antibody (n3kobu5), APC, eBioscience

Applications

• Flow Cytometry (Intracellular staining)

Tested Dilution: 5 µL (0.03 µg)/test
Publications: View 3 publications

Product Specifications

Product Specific Information

This n3kobu5 monoclonal antibody recognizes human and mouse ZAP-70 and SYK when phosphorylated on Y319 and Y352, respectively.
ZAP-70 and SYK are members of the SYK protein tyrosine kinase family activated by Src family kinases.
Phosphorylation of ZAP-70 Y319 by Lck is essential for TCR-dependent signaling, while SYK Y352 phosphorylation by Fyn/Lyn is critical for BCR signaling and B cell development.

Specificity was confirmed by ELISA, flow cytometry, and western blotting.

Applications Reported: Intracellular staining followed by flow cytometric analysis.
Applications Tested: Pre-titrated and validated for intracellular staining of stimulated human peripheral blood cells. Recommended usage: 5 µL (0.03 µg)/test in 100 µL final volume. Cell count may vary from 10⁵ to 10⁸ cells/test.

Staining Protocols

Protocol A: Two-step (cytoplasmic proteins) – optimal for surface and intracellular detection.
Protocol B: One-step (nuclear proteins) – recommended for transcription factor staining.
Protocol C: Two-step (Fixation/Methanol) – best for phospho-specific signaling discrimination.

Detailed protocols available in BestProtocols® - Staining Intracellular Antigens for Flow Cytometry.

Excitation: 633–647 nm
Emission: 660 nm
Laser: Red Laser
Filtration: 0.2 µm post-manufacturing filtered

Target Information

ZAP-70 is a cytosolic protein tyrosine kinase of the Syk family, expressed in T cells and NK cells, essential for TCR activation.
Upon TCR engagement, ZAP70 is phosphorylated on tyrosines 292, 315, and 319.
Phosphorylation of 315 and 319 enhances signaling, while 292 attenuates activation.

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

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