ZAP 70 (TCR Zeta Associated Protein kinase 70kD)

ZAP-70 is a cytosolic protein tyrosine kinase that is a member of the Syk family of proteins. ZAP-70 is expressed exclusively in T cells and natural killer cells and is required for T cell receptor activation, which triggers an immune response. ZAP-70 is activated when the protein tyrosine kinase Lck phosphorylates the tyrosine residues on ZAP-70. Phosphorylation of ZAP-70 is crucial to the recruitment of additional molecules, such as Vav, Ras, and PLC gamma-1, to the membrane-associated signaling complex and for the induction of downstream signaling events. Mutation of the ZAP-70 genes results in a form of Severe Combined Immunode?ciency Syndrome (SCID) in humans. \n\nApplications: \nSuitable for use in Western Blot, Flow Cytometry and Immunoprecipitation. Other applications not tested.\n\nRecommended Dilution: \nWestern Blot: 0.5-2ug/ml. Jurkat cell lysate was resolved by electrophoresis, transferred to nitrocellulose and probed with Z0300-05. Proteins were visualized using a goat anti-mouse secondary antibody conjugated to HRP and a chemiluminescence detection system. \nFlow Cytometry: Detects ZAP-70 in Jurkat cells. 1x10e6 Jurkat cells were stained with 1ug of Z0300-05 followed by a goat anti-mouse conjugated PE antibody.\nImmunoprecipitation: 4ug immunoprecipitated p70 ZAP from 500ug of RIPA lysates from Jurkat or Hut-78 cell lines. \nOptimal dilution determined by the researcher. \n\nIncluded Positive Antigen Control: \nJ9000: Jurkat lysate. Add 2.5ul of 2-mercaptoethanol/100ul of lysate and boil for 5 minutes to reduce the preparation. Load 20ug of reduced lysate per lane for minigels. \n\nStorage and Stability:\nMay be stored at 4 degrees C for short-term only. For long-term storage and to avoid repeated freezing and thawing, add sterile glycerol 40-50%, aliquot and store at -20 degrees C. Aliquots are stable for at least 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

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