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Jurkat cells were either untreated (lane 1) or treated (lane 2) with Anti-CD3 for 15 minutes at 37°C. The top panel was probed with Lck (cat. #610097) and the bottom panel was probed with Lck (pY505) (cat. #612390).
BD Transduction Laboratories™ Purified Mouse Anti-Lck (pY505)
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推荐的实验流程
Western blot: Please refer to http://www.bdbiosciences.com/pharmingen/protocols/Western_Blotting.shtml.
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- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
Protein tyrosine phosphorylation is an essential step in the signal transduction cascade leading to T cell antigen receptor (TCR) activation. Lck is a protein kinase and a member of the src family of cytoplasmic protein-tyrosine kinases (PTKs). Members of this family have several common features: 1) unique N-terminal domains, 2) attachment to cellular membranes through a myristylated N-terminus, and 3) homologous SH2, SH3, and catalytic domains. The unique N-terminal domain of Lck interacts with the cytoplasmic tails of the CD4 and CD8 cell surface glycoproteins. CD4 and CD8 bind to surface MHC class II and class I molecules, respectively. Lck is regulated by both kinases and phosphatases. Autophosphorylation at Y394 leads to conformational changes in the catalytic domain, which induces kinase activity. Repression of Lck occurs via phosphorylation at Y505, located near the carboxy-terminus. Phosphorylation of this tyrosine site is mediated by the Csk family of PTKs. Upon phosphorylation at this site, Lck associates with the SH2 domain in the amino-terminus, thus keeping the protein biologically inactive. Lck activity and regulation is critical for activation and development of T cells.
研发参考 (3)
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Hardwick JS, Sefton BM. The activated form of the Lck tyrosine protein kinase in cells exposed to hydrogen peroxide is phosphorylated at both Tyr-394 and Tyr-505. J Biol Chem. 1997; 272(41):25429-25432. (Biology). 查看参考
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Lee-Fruman KK, Collins TL, Burakoff SJ. Role of the Lck Src homology 2 and 3 domains in protein tyrosine phosphorylation. J Biol Chem. 1996; 271(40):25003-25012. (Biology). 查看参考
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Wang B, Lemay S, Tsai S, Veillette A. SH2 domain-mediated interaction of inhibitory protein tyrosine kinase Csk with protein tyrosine phosphatase-HSCF. Mol Cell Biol. 2001; 21(4):1077-1088. (Biology). 查看参考
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