Biolegend, 683710, PerCP/Cyanine5.5 anti-ZAP70 Phospho (Tyr319)/Syk Phospho (Tyr352) Antibody, 100tests

ZAP70 was identified in TCR-stimulated Jurkat cells. It is an inactive cytosolic tyrosine kinase that is recruited to a transmembrane receptor lacking intrinsic catalytic activity. ZAP70 and the related spleen tyrosine kinase (Syk) play a critical role in T-cell development and activation. This enzyme, which is phosphorylated on tyrosine residues upon T-cell antigen receptor (TCR) stimulation, functions in the initial step of TCR-mediated signal transduction in combination with the Src family kinases, Lck and Fyn. ZAP70 activation can be regulated by binding to phosphorylated ITAMs of the TCR and by phosphorylation of multiple tyrosine residues on ZAP70. Phosphorylation of Tyr-315 and Tyr-319 are essential for ZAP70 positive regulation of T-lymphocyte activation whereas Tyr-292 has a negative regulatory role. Phosphorylated Y319 is a positive regulator of ZAP70, which triggers the binding of Lck and activation of NFAT and IL-2 induction. The binding of Lck to phosphorylated Y319 promotes Lck mediated phosphorylation of Y493, and facilitates activation of downstream signaling. The importance of Y319 phosphorylation in positive regulation of ZAP70 was confirmed in a mouse model with Y319F ZAP70 mutant expression, which resulted in severe defects in calcium mobilization and thymocyte selection.
100tests
Verified Reactivity: Human
Antibody Type: Monoclonal
Host Species: Mouse
Immunogen: Modified peptide.
Formulation: Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide and BSA (origin USA)
Preparation: The antibody was purified by affinity chromatography and conjugated with PerCP/Cyanine5.5 under optimal conditions.
Concentration: Lot-specific (to obtain lot-specific concentration and expiration, please enter the lot number in our Certificate of Analysis online tool.)
Storage & Handling: The antibody solution should be stored undiluted between 2°C and 8°C, and protected from prolonged exposure to light. Do not freeze.
Application: ICFC - Quality tested
Recommended Usage: Each lot of this antibody is quality control tested by intracellular flow cytometry using our True-Phos™ Perm Buffer in Whole Blood Protocol. For flow cytometric staining, the suggested use of this reagent is 5 µL per million cells in 100 µL staining volume or 5 µL per 100 µL of whole blood. It is recommended that the reagent be titrated for optimal performance for each application. * PerCP/Cyanine5.5 has a maximum absorption of 482 nm and a maximum emission of 690 nm.
Excitation Laser: Blue Laser (488 nm)
Application Notes: This antibody can also weakly react with Syk Phospho (Tyr352) on WB.
Product Citations: Gill PS, et al. 2022. Front Immunol. 13:872652. PubMed
RRID: AB_2687050 (BioLegend Cat. No. 683709) AB_2687051 (BioLegend Cat. No. 683710)
Structure: 619 amino acids with a predicted molecular weight of 70 kD.
Distribution: Cytoplasm and cell membrane.
Function: Required for the assembly of a Zap-70-containing signaling complex. Activation of the PLC-g1-dependent and Ras-dependent signaling cascades. Association with PLC-g1.
Interaction: NFAM1, SLA and SLA2, CBLB, DEF6, FCRL3, VAV1, and CD247/CD3Z.
Cell Type: B cells
Biology Area: Cell Biology, Immunology, Signal Transduction
Molecular Family: Phospho-Proteins, Protein Kinases/Phosphatase, TCRs
Antigen References: 1. Chan AC, et al. 1991. Proc. Natl. Acad. Sci. 88:9166. 2. Arpaia E, et al. 1994. Cell 76:947. 3. Chan AC, et al. 1994. Science 264:1599. 4. Negishi I, et al. 1995. Nature 376:435. 5. Wang H, et al. 2010. Cold Spring Harb. Perspect. Biol. 2:a002279. 6. Klammt C, et al. 2015. Nat. Immunol. 16:961. 7. Weiss A. 1993. Cell 73:209. 8. Gong Q, et al. 2001. J. Exp. Med. 194:507.
Gene ID: 7535
UniProt: View information about ZAP-70 Phospho Tyr319 Syk Phospho Tyr352 on UniProt.org
Clone: 1503310
Regulatory Status: RUO
Other Names: Phospho Zeta-Chain (TCR) Associated Protein Kinase 70 kD, Phospho SRK, Phospho 70 KD Zeta-Chain Associated Protein, Phospho Tyrosine-protein kinase ZAP-70
Isotype: Mouse IgG1, κ
Q: How stable is PerCP/Cyanine5.5 tandem as compared to PerCP alone?
A: PerCP/Cyanine5.5 is quite photostable and also better than PerCP alone in withstanding fixation.