Description
The 70-kDa ζ chain-associated protein (ZAP70) is a Syk-family protein tyrosine kinase (PTK) that associates with the ζ subunit of the T cell antigen receptor (TCR) and undergoes tyrosine phosphorylation following TCR stimulation. ZAP70 contains two SH2-like domains with the PTK domain located at the C-terminus. TCR-mediated Lck activity leads to phosphorylation of ZAP70 on Tyrosine 493 in the regulatory loop of the PTK domain leading to upregulation of ZAP70 kinase activity. Tyrosine 292 (Y292), in the linker region between the SH2 and PTK domains, is autophosphorylated by the activated PTK domain. By binding with c-Cbl, the phosphorylated Y292 can down-regulate TCR signaling. The J34-602 antibody recognizes ZAP70 phosphorylated at Y292.
This antibody conjugate is suitable for intracellular staining of human whole blood (using BD Phosflow™ Lyse/Fix Buffer) and peripheral blood mononuclear cells (using BD Cytofix™ Fixation Buffer). Any of the three BD Phosflow™ permeabilization buffers may be used.
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 488 under optimum conditions, and unreacted Alexa Fluor® 488 was removed.
1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
2. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
3. Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
4. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
7. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
8. All other brands are trademarks of their respective owners.
9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
