Vâlcu Cristina-Maria, Schlink Katja
Section of Forest Genetics, Technische Universität München, Freising-Weihenstephan, Germany.
Proteomics. 2006 Mar;6(5):1599-605. doi: 10.1002/pmic.200500314.
Protein extraction procedure and the reducing agent content (DTT, dithioerythritol, tributyl phosphine and tris (2-carboxyethyl) phosphine (TCEP)) of the sample and rehydration buffers were optimised for European beech leaves and roots and Norway spruce needles. Optimal extraction was achieved with 100 mM DTT for leaves and needles and a mixture of 2 mM TCEP and 50 mM DTT for roots. Performing IEF in buffers containing hydroxyethyldisulphide significantly enhanced the quality of separation for all proteins except for acidic root proteins, which were optimally focused in the same buffer as extracted.
针对欧洲山毛榉的叶子和根以及挪威云杉的针叶，优化了蛋白质提取程序、样品和复水缓冲液中的还原剂含量（二硫苏糖醇（DTT）、二硫赤藓糖醇、三丁基膦和三（2-羧乙基）膦（TCEP））。叶子和针叶使用100 mM DTT，根使用2 mM TCEP和50 mM DTT的混合物可实现最佳提取。在含有羟乙基二硫化物的缓冲液中进行IEF，除酸性根蛋白外，所有蛋白质的分离质量均显著提高，酸性根蛋白在提取时所用的相同缓冲液中聚焦效果最佳。
