The T Cell Signaling Antibody Sampler Kit contains primary and secondary antibodies to perform four Western blots with each antibody.
Each antibody in the T Cell Signaling Antibody Sampler Kit recognizes only its specific target and does not cross-react with other family members with the following exceptions: Phospho-Zap-70 (Tyr319)/Syk (Tyr352) (65E4) Rabbit mAb which also recognizes Syk when phosphorylated at Tyr352; and Phospho-Lck (Tyr505) Antibody which may cross-react with certain phosphorylated Src family members.
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding the amino terminus of human Zap-70; Tyr319 of human Zap-70; and Tyr505 of human Lck.
Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the sequence of human SLP-76; residues surrounding Tyr191 of human LAT; the central region of human LAT; and residues surrounding Tyr505 of human Lck. Antibodies are purified by protein A and peptide affinity chromatography.
The Syk family protein tyrosine kinase Zap-70 is expressed in T and NK cells and plays a critical role in mediating T cell activation in response to T cell receptor (TCR) engagement (1). Following TCR engagement, Zap-70 is rapidly phosphorylated on several tyrosine residues through autophosphorylation and transphosphorylation by the Src family tyrosine kinase Lck (2-6). Tyrosine phosphorylation correlates with increased Zap-70 kinase activity and downstream signaling events. Expression of Zap-70 is correlated with disease progression and survival in patients with chronic lymphocytic leukemia (7,8).
LAT, a transmembrane adaptor protein expressed in T, NK and mast cells, is an important mediator for T cell receptor (TCR) signaling (9). Upon TCR engagement, activated Zap-70 phosphorylates LAT at multiple conserved tyrosine residues within SH2 binding motifs, exposing these motifs as the docking sites for downstream signaling targets (10,11). The phosphorylation of LAT at Tyr171 and 191 enables the binding of Grb2, Gads/SLP-76, PLCgamma1 and PI3 kinase through their SH2 domain and translocates them to the membrane. This process eventually leads to activation of the corresponding signaling pathways (11-12).
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc. U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.
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