使用iTRAQ标记和LTQ Orbitrap XL定量分析DNA损伤反应蛋白质

　　简介：To identify and determine relative amounts of DNA damage response proteins in A549 cells over 24 hours after treatment with camptothecin using iTRAQ labeling and the LTQ Orbitrap XL.

　　仪器：LTQ Orbitrap XL

　　结论：

　　Quantitation of iTRAQ-labeled peptides in complex mixtures can be successfully performed using HCD fragmentation. In this study, more than 500 proteins from A549 cell lysates were identified and quantified using iTRAQ labeling and an LTQ Orbitrap XL mass spectrometer equipped with an HCD collision cell. Phosphoprotein purification using the Thermo Scientific Phosphoprotein Enrichment Kit increased sample diversity by adding 145 additional proteins. After sequential IMAC purification of a 15 μg digested PE sample, an additional thirty phosphorylation sites were identified and quantified.