Data-independent proteome analysis of ARPE-19 cells.

Data-independent proteome analysis of ARPE-19 cells.
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DOI:
10.1016/j.dib.2018.06.103
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发表时间:
2018-10
期刊:
影响因子:
1.2
通讯作者:
Giorgianni F
Giorgianni F
中科院分区:
其他
文献类型:
--
作者:
Koirala D;Beranova-Giorgianni S;Giorgianni F

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我们已经对人视网膜色素上皮细胞系(ARPE-19)进行了蛋白质组学分析,其代表了广泛用于体外研究与人RPE细胞相关的细胞和分子机制的模型(Dunn等人,1996; Weigel等人,2002年)。全细胞蛋白质提取物通过短(10分钟)SDS-PAGE运行分离成四个凝胶级分。分级分离和胰蛋白酶消化后,在纳米UPLC LC系统上分离所得肽,并使用QTof-IMS仪器进行在线分析:具有离子迁移率分离的串联质谱仪(Synapt G2-Si)。以数据独立模式(UDMSE)获取数据,其允许绝对和/或相对获取后蛋白质定量(Silva et al.,2006年)。从本研究中获得的蛋白质组谱数据可以用作蛋白质参考数据库,具有与正常生长条件下的ARPE-19细胞相关的定性和定量蛋白质信息。
We have performed a proteomics analysis of a human retinal pigment epithelial cell line (ARPE-19), which represents a widely used model for in vitro studies of cellular and molecular mechanisms related to human RPE cells (Dunn et al., 1996; Weigel et al., 2002). Whole cell protein extracts were separated in four gel fractions via short (10 min) SDS-PAGE runs. Following fractionation and trypsin digestion, the resulting peptides were separated on a nano UPLC LC system and analyzed on-line with a QTof-IMS instrument: a tandem mass spectrometer with ion mobility separation (Synapt G2-Si). Data were acquired in data-independent mode (UDMSE), which allows for absolute and/or relative post-acquisition protein quantification (Silva et al., 2006). The proteome profile data obtained from this study can be used as a protein reference database with qualitative and quantitative protein information related to ARPE-19 cells under normal growth conditions.
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