Evaluation of two-dimensional electrophoresis and liquid chromatography – tandem mass spectrometry for tissue-specific protein profiling of laser-microdissected plant samples

摘要: Laser microdissection (LM) allows the collection of homogeneous tissue- and cell-specific plant samples. The employment this technique with subsequent protein analysis has thus far not been reported for tissues, probably due to difficulties associated defining a reasonable cellular morphology and, in parallel, allowing efficient extraction from tissue relatively large sample amount needed successful proteome is an additional issue that complicates profiling on or even level. In contrast transcript can be performed very small amounts amplification strategies, there as yet no procedure proteins available. current study, we compared different preparation techniques prior LM/laser pressure catapulting (LMPC) respect their suitability retrieval. Cryo-sectioning was identified best compromise between effective extraction. After vascular bundles Arabidopsis thaliana stem by LMPC, were extracted subjected analysis, either classical two-dimensional gel electrophoresis (2-DE), high-efficiency liquid chromatography (LC) conjunction tandem mass spectrometry (MS/MS). Our results demonstrate both methods used LMPC collected material. But because significantly lower required LC-MS/MS than 2-DE, combination higher potential promote comprehensive specific tissues.