二维凝胶电泳和质谱技术.pptVIP

* * MALDI：基质辅助激光解吸电离技术（Matrix-assisted laser desorption ionization) * 极性化合物：溶解样品；芳环：吸收激光；易结晶：均匀分散样品分子常用2，5-二羟基苯甲酸 * Matrix-extraction effect:higher tolerance to impurities 基质提取效应：对不纯样品的高度耐受 Matrix ions:low mass range(500) unusable基质离子:分子量低于500不能采用 Contaminant ions:background peaks污染物离子：背景峰 * The principle here is to have the first quadrupole scan across the mass range that one would expect precursor masses that contain phosphorylation. The precursor ions pass into the second quadrupole region which acts as a collision cell. Gas (usually nitrogen or air which is 80%nitrogen) is introduced and the precursor ions collide with the gas molecules. The fragments exit into the third quadrupole which is tuned to only select a single mass, that of m/z 79 the characteristic marker mass of the phosphate group. The resulting spectrum that is recorded represents only those masses that have given rise to the marker ion of m/z79. All of this is achieved in negative ion mode. In the case of phosphorylated peptides it is way of distinguishing phosphopeptides from normal peptides and effectively “cleans up” the result to only the phosphopeptides. Combined with further MS/MS scan functions this technique is very powerful to both detect and sequence phosphopeptides. * 拥有最新离子源、四极杆、ICR和软件系统。分辨率超过140万，准确度优于0.5ppm，灵敏度attmol，隔离效率优于100%。该系统还配备了丰富的辅助离子解离组件(如ECD，IRMPD)，可实现超高分辨、高准确度和高灵敏度的多级串联质谱功能，可无须酶切而直接鉴定蛋白质，是研究生物标志物最有力的、最可靠的手段。该系统将在“代谢组学”、“中药现代化”、“蛋白质组学”,“临床蛋白质组学”、“多肽组学”、“免疫组学”、“生命环境科学”等多个领域开展深层次研究。 MALDI AND ESI MALDI 固相 高能离子轰击 单电荷离子 基质离子 易操作 ESI 液相 强电场 多电荷离子 污染离子 操作相对复杂 质量分析器 Mass analysis 质量分析器是质谱仪的核心,它将离子源产生的离子按其质荷比(m/z) 的不同﹑在空间的位置﹑时间的先后或轨道的稳定与否进行分离,以便得到按m/z大小顺序排列而成的质谱图。 两组四极杆分别加上 +(u+Vcon?t) 和 –(u+Vcon?t)， 其中u直流电压部分，Vcon?t为射频电压部分 ? = 2?f, f为频率。 一、四极杆质量分析器(Quadrupole Mass Analyser) 在场的中心和两条对角线上的任何一点电位为零。 通过直流和射频电压的改变来控制不同荷质比的离子通过； 可以允许单一荷质比的离子通过；也可以进行荷质比扫描 mass scanning mode m1 m3 m4 m2 m3 m1 m4 m2 single mass transmission mode m2 m2 m2 m2 m3 m1 m4 m2 四级杆扫描模式 离子在四级杆中的稳定性 三重四级杆 离子阱质