Syk is a non-receptor protein-tyrosine kinase that is closely related to ZAP70 and plays crucial roles in the development and receptor-mediated signaling of most leukocytes and in vascular integrity. Syk is expressed in hematopoietic cells, including B lymphocytes, immature (CD4, CD8 double-negative and double-positive) thymocytes, and myeloid cells, epithelial cell lines, and normal breast tissue. Mature (CD4 or CD8 single-positive) thymocytes and peripheral αβ TCR-bearing T lymphocytes have very low or undetectable levels of Syk. Syk contributes to the signal transduction process by binding to ITAMs (Immunoreceptor Tyrosine-based Activation Motifs) of immune receptors, including Igα and Igβ (CD79a and b), TCRζ, CD3ε, and FcRγ. Upon receptor activation, Syk binds to phosphorylated ITAMs via its two N-terminal SH2 domains thereby activating Syk and causing tyrosines in the interdomain, between the SH2 and Kinase domains of Syk, to undergo auto-phosphorylation and phosphorylation by Lyn. The tyrosine 348 phosphorylation site (pY348) in human Syk is orthologous to tyrosine 342 in mouse and rat Syk and tyrosine 315 in human ZAP70. This phosphorylated site can act as a binding site for other signaling molecules, such as PLCγ, Vav, and Fgr. The I120-772 antibody is specific for human Syk (pY348) and does not cross-react with phosphorylated Zap70. The orthologous phosphorylation site in mouse and rat Syk is Y342. Analysis of Syk (pY348) in human peripheral blood lymphocytes. Human peripheral blood mononuclear cells (PBMC) were either stimulated (right panel) by cross-linking of surface IgM with purified Goat F(ab')2 Anti-Human IgM (SouthernBiotech) at 37 °C for 8 minutes or unstimulated (left panel). The cells were fixed with pre-warmed BD Phosflow™ Lyse/Fix Buffer (Cat. No. 558049) for 10 minutes at 37 °C, permeabilized with BD Phosflow™ Perm Buffer II (Cat. No. 558052) at room temperature for 10 minutes, and then stained with PE Mouse anti-Syk (pY348) and Alexa Fluor® 647 Mouse anti-human CD20 mAb H1(FB1) (Cat. No. 558054). For data analysis, lymphocytes were selected by their scatter profile. The data demonstrates that the upregulated phosphorylation is restricted to the stimulated CD20-positive B lymphocytes. Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.
BD Phosflow™ PE Mouse anti-Syk (pY348) - PE - Clone I120-722 - Isotype Mouse IgG1, κ - Reactivity Hu, Ms - 50 Tests
Purification
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
SYK
Reactivité: Humain
WB, IHC, ELISA, IF
Hôte: Lapin
Polyclonal
unconjugated
Indications d'application
Optimal working dilution should be determined by the investigator.
Volume d'échantillon
20 μL
Restrictions
For Research Use only
Buffer
Aqueous buffered solution containing BSA and ≤0.09 % sodium azide.
Agent conservateur
Sodium azide
Précaution d'utilisation
This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Conseil sur la manipulation
The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
Stock
4 °C
Stockage commentaire
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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