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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Cell cycle checkpoints are regulatory mechanisms that prevent cell cycle progression in the presence of DNA damage or incompletely replicated DNA. Many of the genes required for cell-cycle arrest are also involved in DNA repair, apoptosis, and induction of transcription. In yeast and humans, hRAD9 plays a role in cell cycle arrest during the G2 phase before entry into mitosis. Phosphorylated hRAD9 is found in the nucleus after DNA damage, and forms DNA damage-responsive complexes with other putative checkpoint control proteins, such as hRAD1 and hHUS1. Expression of hRAD9 in S. pombe rad9::ura4+ cells restores resistance to the DNA synthesis inhibitor hydroxyurea and gamma rays. In addition, hRAD9 binds the anti-apoptotic proteins, Bcl-2 and Bcl-xL , and antisense hRAD9 RNA suppresses DNA-damage induced cell death. Thus, hRAD9 may be an important component of protein complexes that regulate cell cycle progression, as well as apoptosis, in response to DNA damage.
研发参考 (5)
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Komatsu K, Miyashita T, Hang H. Human homologue of S. pombe Rad9 interacts with BCL-2/BCL-xL and promotes apoptosis. Nat Cell Biol. 2000; 2(1):1-6. (Biology). 查看参考
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Lieberman HB, Hopkins KM, Nass M, Demetrick D, Davey S. A human homolog of the Schizosaccharomyces pombe rad9+ checkpoint control gene. Proc Natl Acad Sci U S A. 1996; 93(24):13890-13895. (Biology). 查看参考
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St Onge RP, Udell CM, Casselman R, Davey S. The human G2 checkpoint control protein hRAD9 is a nuclear phosphoprotein that forms complexes with hRAD1 and hHUS1. Mol Cell Biol. 1999; 10(6):1985-1995. (Biology). 查看参考
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Volkmer E, Karnitz LM. Human homologs of Schizosaccharomyces pombe rad1, hus1, and rad9 form a DNA damage-responsive protein complex. J Biol Chem. 1999; 274(2):567-570. (Biology). 查看参考
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Yoshida K, Komatsu K, Wang HG, Kufe D. c-Abl tyrosine kinase regulates the human Rad9 checkpoint protein in response to DNA damage. Mol Cell Biol. 2002; 22(10):3292-32300. (Biology: Immunoprecipitation, Western blot). 查看参考
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