DIGE荧光差异双向电泳.pdfVIP

2D-DIGE 荧光差异双向电泳 Why 2D-DIGE？ 2 November 2012 当传统双向电泳面临新的要求时……  重复性  定量精确性  灵敏度  线性动态范围  质谱兼容与否 2-D DIGE - 精准的定量技术，检测到更多低丰度蛋白 Total number of DIGE publications ~1500 publications Top 5 categories of DIGE publications: 1. Human medicine 2. Proteomics 3. Molecular biology 4. Plants 5. Environment 传统双向电泳方法 Control Rat 1 Control Rat 2 Control Rat 3 Control Rat 4 24 Gels Treated Rat 1 Treated Rat 2 Treated Rat 3 Treated Rat 4 2-D DIGE 节约时间 减少6倍工作 Control Rat 1 Control Rat 2 Control Rat 3 Control Rat 4 Treated Rat 1 Treated Rat 2 Treated Rat 3 Treated Rat 4 胶间重复：每组每只大鼠的样本要重复三次，降低系统误差 生物重复：每组要有不同大鼠样本的重复，排除生物学差异 传统2D：2 ×3 ×4 ＝24 gels DIGE：2 ×4/2=4 gels 2-D DIGE 定量结果更准确 消除实验中系统误差 Gel to gel variation Large gel-to-gel variation Small gel-to-gel variation 6 replicate gels made from the same sample (SYPRO™ Ruby) Data courtesy of Jörgen