The relationship between protein concentration of lysates from untreated and H2O2-treated Jurkat cells and the absorbance at 450 nm using PathScan® Phospho- Zap-70 (Tyr317) Sandwich ELISA Antibody Pair #7852 is shown. Jurkat cells (0.8 x 106 cells/ml) were treated with H2O2 (2 mM) for 2 min at 25ºC and then lysed.
Capture and detection antibodies are stored at 4°C. HRP-linked secondary reagent is stored at -20°C.
CST's PathScan® Phospho-Zap-70 (Tyr317) Sandwich ELISA Antibody Pair is being offered as an economical alternative to our PathScan® Phospho-Zap-70 (Tyr317) Sandwich ELISA Kit #7171. Capture and detection antibodies (100X stocks) and HRP-linked secondary antibody (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The Zap-70 capture antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysates are added followed by a Phospho-Zap-70 (Tyr317) detection antibody and anti-Rabbit IgG, HRP-linked antibody. HRP substrate TMB is added for color development. The magnitude of the absorbance for this developed color is proportional to the quantity of phospho- Zap-70 (Tyr317) protein.
For Antibody Pair specificity and sensitivity, please refer to the corresponding PathScan® Sandwich ELISA Kit. Note: This antibody pair detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
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).
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PathScan is a trademark of Cell Signaling Technology, Inc.
U.S. Patent No. 7,429,487, foreign equivalents, and child patents deriving therefrom.