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Western blot analysis of SHC on a HeLa cell lysate (Human cervical epitheloid carcinoma; ATCC CCL-2.2). Lane 1: 1:1000, lane 2: 1:2000, lane 3: 1:4000 dilution of the Mouse Anti-SHC antibody.
Immunofluorescence staining of WI-38 cells (Human lung fibroblasts; ATCC CCL-75).
BD Transduction Laboratories™ Purified Mouse Anti-SHC
BD Transduction Laboratories™ Purified Mouse Anti-SHC
监管状态图例
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准备和存储
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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
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The mammalian SHC proteins, which are expressed as multiple isoforms (46, 52, and 66 kDa), each contain a C-terminal SH2 domain and an N-terminal glycine/proline rich sequence. These proteins function as early signaling intermediates (also called adaptor proteins) which relay G protein coupled receptor (GPCR) and receptor tyrosine kinase (RTK)-induced signals via the Ras transduction pathway. To this end, the SHC proteins contain specific tyrosine residues which are phosphorylated following association with the active RTKs. Phosphorylated SHC forms a complex with the adaptor protein GRB2. Association of the SHC-GRB2 complex with the Ras guanine nucleotide exchange factor (Ras-GEF) mediates the localization of Ras-GEF to the plasma membrane. Once at the plasma membrane, Ras-GEF activates Ras by catalyzing the Ras-GTP for Ras-GDP exchange. Over-expression of SHC results in cell transformation, and phosphorylation of SHC correlates with activation of the ERK1/ERK2 kinases. The SHC proteins are mediators of signals that are essential for cell metabolism, growth, and differentiation.
研发参考 (5)
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Imokawa G, Kobayasi T, Miyagishi M. Intracellular signaling mechanisms leading to synergistic effects of endothelin-1 and stem cell factor on proliferation of cultured human melanocytes. Cross-talk via trans-activation of the tyrosine kinase c-kit receptor. J Biol Chem. 2000; 275(43):33321-33328. (Biology: Immunoprecipitation, Western blot). 查看参考
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Kiely PA, Sant A, O'Connor R. RACK1 is an insulin-like growth factor 1 (IGF-1) receptor-interacting protein that can regulate IGF-1-mediated Akt activation and protection from cell death. J Biol Chem. 2002; 277(25):22581-22589. (Biology: Immunoprecipitation, Western blot). 查看参考
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Laser M, Willey CD, Jiang W. Integrin activation and focal complex formation in cardiac hypertrophy. J Cell Biol. 2000; 275(45):35624-35630. (Biology: Western blot). 查看参考
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Pelicci G, Lanfrancone L, Grignani F. A novel transforming protein (SHC) with an SH2 domain is implicated in mitogenic signal transduction. Cell. 1992; 70(1):93-104. (Biology). 查看参考
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Ugi S, Imamura T, Ricketts W, Olefsky JM. Protein phosphatase 2A forms a molecular complex with Shc and regulates Shc tyrosine phosphorylation and downstream mitogenic signaling. Mol Cell Biol. 2002; 22(7):2375-2387. (Biology: Immunoprecipitation, Western blot). 查看参考
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