Biolegend, 693804, PE anti-ZAP70 Phospho (Tyr292) 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 Tyr315 and Tyr319 are essential for ZAP70 positive regulation of T-lymphocyte activation whereas Tyr292 has a negative regulatory role. Mutation of Tyr292 to phenylalanine enhanced NFAT induction in response to BCR stimulation, indicating phosphorylation of Tyr292 may promote association of ZAP-70 with a negative regulatory protein. ZAP-70 has been reported to bind Cbl following TCR stimulation and phosphorylation of Tyr292 was required for this interaction, as well as for the negative effect of Cbl on signaling.
100tests
Verified Reactivity: Human
Antibody Type: Monoclonal
Host Species: Mouse
Immunogen: Human ZAP70 peptide phosphorylated at Tyr292. Complete Freund's adjuvant.
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 PE 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.
Excitation Laser: Blue Laser (488 nm)Green Laser (532 nm)/Yellow-Green Laser (561 nm)
Application Notes: During development, reactivity was observed with mouse phospho-SYK.
RRID: AB_2686946 (BioLegend Cat. No. 693803) AB_2686947 (BioLegend Cat. No. 693804)
Structure: 619 amino acids with a predicted molecular weight of 70 kD.
Distribution: Cytoplasm; Human T Cells, Mouse T cells and B cells.
Function: Required for the assembly of a Zap-70-containing signaling complex. Activation of the PLC-γ1-dependent and Ras-dependent signaling cascades. Associates with PLC-γ 1.
Interaction: NFAM1, Cbl, SLA and SLA2, CBLB, DEF6, FCRL3, VAV1, CD247/CD3Z.
Cell Type: B cells, T cells
Biology Area: Cell Biology, Immunology
Molecular Family: Phospho-Proteins, Protein Kinases/Phosphatase, TCRs
Antigen References: 1. Chan AC, et al. 1991. Proc. Natl. Acad. Sci. USA 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, et al. 1993. Cell 73:209. 8. Gong Q, et al. 2001. J. Exp. Med. 194:507. 9. Magnan A, et al. 2001. J. Exp. Med. 194:491.
Gene ID: 7535
UniProt: View information about ZAP70 Phospho Tyr292 on UniProt.org
Clone: A16038B
Regulatory Status: RUO
Other Names: p Zeta-Chain (TCR) Associated Protein Kinase 70 kD, p 70 KD Zeta-Chain Associated Protein, p Tyrosine-protein kinase ZAP-70
Isotype: Mouse IgG1, κ
Q: What type of PE do you use in your conjugates?
A: We use R-PE in our conjugates.