高迁移率族蛋白(high mobility group protein,HMG)是一系列的染色质相关蛋白,广泛存在于真核生物细胞中,含量丰富,因其在聚丙烯酰胺凝胶电泳中的高迁移率而得名.HMG蛋白家族可分为HMGB、HMGA和HMGN三类亚家族,各亚家族有其特征的结构域...高迁移率族蛋白(high mobility group protein,HMG)是一系列的染色质相关蛋白,广泛存在于真核生物细胞中,含量丰富,因其在聚丙烯酰胺凝胶电泳中的高迁移率而得名.HMG蛋白家族可分为HMGB、HMGA和HMGN三类亚家族,各亚家族有其特征的结构域,这些结构域介导了HMG和DNA或染色质相关区域的相互作用.现已发现这些蛋白质具有多种重要生物学功能,其中几乎所有HMG都可以通过修饰、弯曲或改变染色质/DNA的结构,促进各种蛋白质因子形成大分子复合物来调节基因转录.展开更多
目的:通过对影响细胞ELISA的关键条件和步骤进行优化和改进,以降低实验假阳性率,提高敏感性。方法:THP-1细胞接种96孔培养板,分别用脂多糖(LPS)刺激或不刺激作为处理组和对照组,将通过噬菌体展示技术筛选获得的噬菌体分别与处理组及对...目的:通过对影响细胞ELISA的关键条件和步骤进行优化和改进,以降低实验假阳性率,提高敏感性。方法:THP-1细胞接种96孔培养板,分别用脂多糖(LPS)刺激或不刺激作为处理组和对照组,将通过噬菌体展示技术筛选获得的噬菌体分别与处理组及对照组细胞作用,洗脱未结合的噬菌体,加入HRP标记的抗M13单克隆抗体,分别用酶标仪和Kodak IS 2000R数码成像系统检测细胞与短肽的结合情况。并针对细胞的特性,在细胞的准备、固定、孵育和洗涤等关键步骤进行改进。结果:酶标仪和数码成像系统检测分别得到6个和8个可与细胞高亲和力结合的阳性噬菌体克隆,经过免疫荧光实验证实数码成像系统检测得到的8个阳性克隆均与细胞结合,并且数码成像系统多次测量值具有很好的重复性。结论:本方法为蛋白质与完整细胞的亲和活性研究提供了有效的实验方案,具有较好的应用前景。展开更多
Sepsis is systemic inflammatory response syndrome(SIRS)associated with the presence of pathogenic microorganisms or their toxin.Lipopolysaccharide(LPS)is an important component of the outer membrane of Gram negative b...Sepsis is systemic inflammatory response syndrome(SIRS)associated with the presence of pathogenic microorganisms or their toxin.Lipopolysaccharide(LPS)is an important component of the outer membrane of Gram negative bacteria and has a pivotal role in inducing Gram negative sepsis.Macrophages play an essential role in infection and inflammation.Specific recognition of LPS is of particular importance in the defense system and pattern recognition receptors(PRR)have been considered to be important in initial steps for cellular recognition of LPS and consequence initiation of LPS responses.In the past few years,intense research in the fields of PRR and their recognition mechanism has been achieved.In this review,we attempt to expound recent research advances of macrophage PRRs for LPS recognition and their mechanism.展开更多
文摘高迁移率族蛋白(high mobility group protein,HMG)是一系列的染色质相关蛋白,广泛存在于真核生物细胞中,含量丰富,因其在聚丙烯酰胺凝胶电泳中的高迁移率而得名.HMG蛋白家族可分为HMGB、HMGA和HMGN三类亚家族,各亚家族有其特征的结构域,这些结构域介导了HMG和DNA或染色质相关区域的相互作用.现已发现这些蛋白质具有多种重要生物学功能,其中几乎所有HMG都可以通过修饰、弯曲或改变染色质/DNA的结构,促进各种蛋白质因子形成大分子复合物来调节基因转录.
文摘目的:通过对影响细胞ELISA的关键条件和步骤进行优化和改进,以降低实验假阳性率,提高敏感性。方法:THP-1细胞接种96孔培养板,分别用脂多糖(LPS)刺激或不刺激作为处理组和对照组,将通过噬菌体展示技术筛选获得的噬菌体分别与处理组及对照组细胞作用,洗脱未结合的噬菌体,加入HRP标记的抗M13单克隆抗体,分别用酶标仪和Kodak IS 2000R数码成像系统检测细胞与短肽的结合情况。并针对细胞的特性,在细胞的准备、固定、孵育和洗涤等关键步骤进行改进。结果:酶标仪和数码成像系统检测分别得到6个和8个可与细胞高亲和力结合的阳性噬菌体克隆,经过免疫荧光实验证实数码成像系统检测得到的8个阳性克隆均与细胞结合,并且数码成像系统多次测量值具有很好的重复性。结论:本方法为蛋白质与完整细胞的亲和活性研究提供了有效的实验方案,具有较好的应用前景。
文摘Sepsis is systemic inflammatory response syndrome(SIRS)associated with the presence of pathogenic microorganisms or their toxin.Lipopolysaccharide(LPS)is an important component of the outer membrane of Gram negative bacteria and has a pivotal role in inducing Gram negative sepsis.Macrophages play an essential role in infection and inflammation.Specific recognition of LPS is of particular importance in the defense system and pattern recognition receptors(PRR)have been considered to be important in initial steps for cellular recognition of LPS and consequence initiation of LPS responses.In the past few years,intense research in the fields of PRR and their recognition mechanism has been achieved.In this review,we attempt to expound recent research advances of macrophage PRRs for LPS recognition and their mechanism.
