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PRM(平行反应监视,parallel reaction monitoring) is一种based on高分辨, 高精度质谱of离子监视技术, is对传统质谱定量技术SRM/MRMof升级版, 该技术be able to对目标protein, 目标肽段 (such as发生翻译后修饰of肽段) 进行选择性detect, 从而实现对目标protein/肽段进行绝对定量.中选生物翻译后修饰PRM靶向validate组学服务provide磷酸化, 乙酰化, 泛素化etc.翻译后修饰后PRM靶向validate, 可代替Western Blot.
技术特色:
(1) 质量精度达到ppm级, be able to比SRM/MRM更好地排除背景干扰and假阳性, have效提高复杂背景下ofDetection LimitandSensitivity;
(2) 对子离子进行全扫描, 无需选择离子对andoptimize碎裂能量, 更容易进行方法学建立;
(3) 更宽of线性范围: 增加至5-6pcs数量级;
(4) 可替代western blotvalidate.
Sample Type | 样本要求 |
动物tissue | ≥100mg |
植物tissue | ≥500mg |
cell样品 | ≥2x10^7
|
血液etc.体液 | ≥1ml |
microorganism类 (菌类) | ≥300mg |
目标修饰位点of选择: 选择合适of修饰位点进行validateisPRM实验of关键. 由于修饰usually会导致proteinof结构and性质发生变化, 因此need选择那些对修饰敏感且具have代表性of位点.
内标准of选择with制备: 对于修饰protein, 内标准of选择and制备可能more复杂. usually, 内标准shouldiswith目标修饰protein结构相似但未被修饰ofprotein, oris具have相同修饰但修饰程度不同ofprotein.
修饰程度of定量: 修饰proteinusually存in多种修饰程度, such as何accurate定量每种修饰程度is一pcs挑战. 这可能needusing多pcs目标肽段and相应of内标准, as well as精确ofData ProcessingandAnalysis Method.
质谱条件ofoptimize: for了获得most佳ofPRM结果, 可能need对质谱条件进行optimize, including肽段of选择, 碰撞能量of调整, 质谱分辨率of设定etc.
