Preparation And Storage
Recommended Assay Procedures
Western blot: Please refer to http://www.bdbiosciences.com/pharmingen/protocols/Western_Blotting.shtml
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
ZAP-70 is a protein tyrosine kinase (PTK) that associates with the ζ subunit of the T cell antigen receptor (TCR) and undergoes tyrosine phosphorylation following TCR stimulation. ZAP-70 contains two SH2-like domains with the PTK domain being at the C-terminus. It appears that both ZAP-70 and Syk are recruited to the phosphorylated CD3 and ζ subunits after TCR stimulation. The src-family PTKs, lck, and fyn, seem to play a role upstream of ZAP-70 and Syk where they phosphorylate CD3 and ζ. Thus both Src- and ZAP-70/Syk-family PTKs are critical for efficient induction of tyrosine phosphoproteins in heterologous cells. The significance of ZAP-70 in mediating TCR signal transduction has been confirmed by showing that ZAP-70 activity is absent in an autosomal recessive form of severe combined immunodeficiency (SCID). This is due to mutations affecting the ZAP-70 kinase domain which affect the stability of the protein. The levels of lck and fyn appear to be normal in the same cells. Results of the ZAP-70 mutations in these patients include an unusual absence of peripheral CD8+ T cells and an abundance of CD4+ T cells in peripheral lymphoid organs. These results indicate that these two populations of T cells develop through utilization of different intracellular signaling pathways.
Development References (5)
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Arpaia E, Shahar M, Dadi H, Cohen A, Roifman CM. Defective T cell receptor signaling and CD8+ thymic selection in humans lacking zap-70 kinase. Cell. 1994; 76(5):947-958. (Biology). View Reference
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Chan AC, Kadlecek TA, Elder ME, et al. ZAP-70 deficiency in an autosomal recessive form of severe combined immunodeficiency. Science. 1994; 264:1599-1601. (Biology).
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Maccalli C, Pisarra P, Vegetti C, Sensi M, Parmiani G, Anichini A. Differential loss of T cell signaling molecules in metastatic melanoma patients' T lymphocyte subsets expressing distinct TCR variable regions. J Immunol. 1999; 163(12):6912-6923. (Biology: Flow cytometry). View Reference
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Michel F, Attal-Bonnefoy G, Mangino G, Mise-Omata S, Acuto O. CD28 as a molecular amplifier extending TCR ligation and signaling capabilities. Immunity. 2001; 15(6):935-945. (Biology: Western blot). View Reference
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Mora AL, Stanley S, Armistead W, Chan AC, Boothby M. Inefficient ZAP-70 phosphorylation and decreased thymic selection in vivo result from inhibition of NF-kappaB/Rel. J Immunol. 2001; 167(10):5628-5635. (Biology: Western blot). View Reference
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