Description: The 1E7.2 antibody reacts with human and mouse ZAP-70, the TCR?-associated protein-70. ZAP-70 is a cytosolic protein tyrosine kinase (PTK) and a member of the Syk family of proteins. It is expressed in T and NK cells and is required for TCR signaling and development. ZAP-70 interacts with the TCR complex by binding to tyrosine-phosphorylated immunoreceptor tyrosine-based activation motifs (ITAMs) present in the invariant subunits of the TCR complex. Following activation, ZAP-70 is phosphorylated on several tyrosine residues by two mechanisms; an autophosphorylation and a transphosphorylation by the Src family tyrosine kinase Lck1-3. Tyrosine phosphorylation of ZAP-70 correlates to its increased kinase activity and triggers downstream signaling events. Mutations in ZAP-70 have been shown to result in a form of Severe Combined Immunodeficiency Syndrome (SCID) in humans. 1E7.2 was generated against a KLH-peptide sequence corresponding to the human ZAP-70 amino acid residues 282-307. While ZAP-70 is normally expressed in T and NK cells, several recent studies have also shown high correlation of ZAP-70 positive expression with mutated IgVH expression in B-chronic lymphocytic leukemia (CCL). In conclusion, the expression of ZAP-70, which can be measured by intracellular flow cytometry, may serve as a prognostic marker for B-CLL. Applications Reported: The 1E7.2 antibody has been reported for use in intracellular flow cytometric analysis.Applications Tested: This 1E7.2 antibody is offered in 2 formats: - µg size: has been tested by intracellular flow cytometric analysis of mouse thymocytes and human Jurkat cells using the Foxp3/Transcription Factor Buffer Set (cat. 00-5523) and protocol. Please refer to Best Protocols: Protocol B: One step protocol for (nuclear) intracellular proteins. This can be used at less than or equal to 1 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. - test size: has been pre-titrated and tested by intracellular flow cytometric analysis of mouse thymocytes and human Jurkat cells using the Foxp3/Transcription Factor Buffer Set (cat. 00-5523) and protocol. Please refer to Best Protocols: Protocol B: One step protocol for (nuclear) intracellular proteins. This can be used at 5 µL (0.5 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. Excitation: 488-561 nm; Emission: 578 nm; Laser: Blue Laser, Green Laser, Yellow-Green Laser. Filtration: 0.2 µm post-manufacturing filtered. ZAP70, a 70 kDa member of the Syk tyrosine kinase family, plays a central role in lymphocyte activation and development, and is implicated in several immune disorders. Upon T cell antigen receptor (TCR) engagement, ZAP70 is phosphorylated on tyrosines 292, 315 and 319 in the interdomain B, located between the SH2 and kinase domains. Phosphorylation of both tyrosines 315 (a Vav binding site) and 319 (a Lck binding site) enhances ZAP70 function in mediating lymphocyte signaling, while tyrosine 292 terminates the transient activation of ZAP70 and attentuates lymphocyte signaling.
|4° C, store in dark, DO NOT FREEZE!|
|PBS with 0.1% gelatin and 0.09% sodium azide; pH 7.2|
For Research Use Only.
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