Anti-Phospho-Zap70-Y319+Syk-Y352 Rabbit Polyclonal Antibody for WB, IF/ICC, ELISA - P43403/P43405

Core Product Specifications and Parameters

Biological Background: ZAP70 Function and Localization

• ZAP70 (70 kDa zeta-chain associated protein) is a cytoplasmic tyrosine kinase belonging to the Syk family, also known as Syk-related tyrosine kinase (SRK).
• Essential for T-cell receptor (TCR) signaling: upon TCR engagement, ZAP70 is recruited to phosphorylated ITAM motifs on CD3 chains, where it is phosphorylated and activated by LCK, initiating downstream signaling cascades. Related references: PMID:7509083
• Activated ZAP70 phosphorylates adaptor proteins LAT and LCP2, triggering a signaling network that regulates T-cell activation, proliferation, cytokine production, and differentiation.
• In addition to TCR signaling, ZAP70 controls T-cell adhesion, motility, and cytoskeletal reorganization; it also mediates TCR internalization and degradation via interaction with CBL, SLA, and SLA2, thereby modulating surface TCR levels.
• ZAP70 also contributes to B-cell development and activation through B-cell receptor (BCR) signaling.
• During thymocyte development, ZAP70 promotes survival and cell-cycle progression of CD4/CD8 double-negative thymocytes prior to positive selection.
• Subcellular localization: predominantly cytoplasmic, but translocates to the plasma membrane upon TCR stimulation.
• Tissue expression: mainly in T-cells and natural killer cells, with lower levels in early thymocytes and pro/pre B-cells.
• Key features include N-terminal tandem SH2 domains that bind phosphorylated ITAMs, a kinase domain with an ATP-binding site, and multiple phosphorylation sites (e.g., Y319) that regulate activity; annotated with keywords such as phosphoprotein, alternative splicing, and disease variant.

Experimental Guidance and Technical Tips

• The immunogen is a synthetic phosphopeptide around Y319 of human ZAP70; the antibody also recognizes the corresponding phospho-Y352 of Syk due to sequence similarity around the phosphorylation site.
• For western blotting, consider using lysates from T-cell lines stimulated with anti-CD3 or pervanadate to induce robust phosphorylation; include a dephosphorylation control (e.g., λ-phosphatase treatment) to confirm phospho-specificity.
• In immunofluorescence/ICC, optimize fixation and permeabilization conditions to preserve both the phospho-epitope and cellular architecture; counterstain with a total ZAP70 antibody to assess activation status.
• For ELISA, a phospho-peptide coating antigen may be employed; ensure blocking steps minimize non-specific binding.
• Cross-reactivity with mouse allows detection in murine T-cell models; however, validate reactivity in the specific experimental system.
• The predicted molecular weight of ZAP70 is ~70 kDa; for Syk, detect the ~72 kDa isoform. In WB, this may appear as closely spaced bands or a single band depending on resolution.

CamelBio: Your One-Stop Sourcing Bridge

As a one-stop IVD raw-material sourcing partner, CamelBio provides diagnostic manufacturers and research laboratories with dependable antibody pairs, bulk ancillary reagents, and tailored procurement solutions for immune signaling targets. This phospho-Zap70/Syk antibody supports investigations into T-cell and B-cell activation pathways, and we offer complementary total protein antibodies, blocking buffers, and rare target sourcing to streamline your assay development from concept to clinic.