蛋白质样品制备技术ppt课件

Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University 1.学习和掌握蛋白质含量测定的原理2.掌握考马斯亮蓝染色法测定蛋白质浓度的操作技术 Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University 准确灵敏：BCA试剂的蛋白质测定范围是202000g/ml；Micro BCA试剂测定范围是0.5-20g/ml快速：45分钟内完成测定，比经典的Lowry法快4倍而且更加方便经济实用：除试管外，测定可在微孔板中进行，大大节约样品和试剂用量不受样品中离子型和非离子型去污剂影响检测不同蛋白质分子的变异系数小于考马斯亮蓝总蛋白定量分析总蛋白定量分析BCA（Bicinchoninic acid，二辛可宁酸、二羧基二喹啉），二辛可宁酸、二羧基二喹啉）Bioinformatics,2010-2011,Shanxi Agricultural University 基本原理：基本原理：碱性条件下，蛋白将Cu2+还原为Cu+，Cu+与BCA试剂形成紫颜色的络合物，测定其在562nm处的吸收值，并与标准曲线对比，即可计算待测蛋白的浓度。Bioinformatics,2010-2011,Shanxi Agricultural University BCA蛋白质检测流程：蛋白质检测流程：Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Coomassie Dye-Based 蛋白质定量蛋白质定量PROTEIN+Amax=595nmBLUEAcidCoomassie G-250Protein-DyeComplexO CH2CH3NHCCH3CH3NCH2SO3-CH3CH2NCH2CH2CH3SO3Na+Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University Bioinformatics,2010-2011,Shanxi Agricultural University 1.下列哪种方法是由于细胞内冰晶的形成导致细胞破碎的（）A.真空干燥 B.酶解法 C.冻融法 D.渗透压法2.当向蛋白质纯溶液中加入中性盐时，蛋白质溶解度（）A增大 B.减小 C.先增大，后减小 D.先减小，后增大3.盐析法与有机溶剂沉淀法比较，其优点是（）A.分辨率高 B.变性作用小 C.杂质易除 D.沉淀易分离4.蛋白质的翻译后修饰主要包括 。A 乙酰化 B 糖基化 C 磷酸化 D 上述各种修饰5.Triton X-100是一种：()A Ca螯合剂 B 蛋白酶抑制剂 C 去垢剂 D 脱氢酶抑制剂Bioinformatics,2010-2011,Shanxi Agricultural University 8、哪一种蛋白质组分在280nm处，具有较大的光吸收？（B ）（中国海洋大学2007）A 色氨酸吲哚基 B 酪氨酸苯酚基 C 苯丙氨酸苯环 D 半胱氨酸的巯基 E 肽链中的肽键9、蛋白质在280nm波长处有特征吸收峰，最大贡献的氨基酸残基是哪一种：（）（厦门大学2006试题）（A）精氨酸（B）组氨酸（C）苯丙氨酸（D）色氨酸