Toward an integrated microchip sized 2-D polyacrylamide slab gel electrophoresis device for proteomic analysis
Toward an integrated microchip sized 2-D polyacrylamide slab gel electrophoresis device for proteomic analysis
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DOI：
10.1002/elps.200600334
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发表时间：
2007-02-01
期刊：
影响因子：
2.9
通讯作者：
Baumann, Marc
中科院分区：
文献类型：
--
作者：
Demianova, Zuzana;Shimmo, Masahiko;Baumann, Marc
我们描述了一个小型化的仪器能够进行2-DE。我们的小型化设备能够在同一单元中进行IEF和聚丙烯酰胺平板凝胶电泳（PASGE）。它由一个用于第一维IEF凝胶的隔室组成，该隔室连接到第二维PASGE凝胶。聚焦的样品通过电迁移自动从IEF凝胶转移到PASGE凝胶。我们的初步实验表明，该设备能够在大约1小时内聚焦和分离蛋白质混合物，不包括染色程序所需的时间。平均而言，凝胶间阻滞因子（Rf）变异为6.2%（+/- 0.9%），p/变异为2.5%（+/- 0.6%）。从染色的凝胶上切下分离的蛋白质点，用胰蛋白酶消化，并通过MS进一步鉴定，从而能够对分离的蛋白质进行直接蛋白质组学分析。
We describe a miniaturized instrument capable of performing 2-DE. Our miniaturized device is able to perform IEF and polyacrylamide slab gel electrophoresis (PASGE) in the same unit. It consists of a compartment for a first-dimensional IEF gel, which is connected to a second-dimensional PASGE gel. The focused samples are automatically transferred from the IEF gel to the PASGE gel by electromigration. Our preliminary experiments show that the device is able to focus and separate a mixture of proteins in approximately 1 h, excluding the time required for the staining procedure. On average, the gel-to-gel retardation factor (Rf) variation was 6.2% (+/- 0.9%) and p/variation was 2.5% (+/- 0.6%). Separated protein spots were excised from stained gels, digested with trypsin, and further identified by MS, thus enabling direct proteomic analysis of the separated proteins.