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从培养的细胞中同时提取蛋白质和代谢物。

Simultaneous extraction of proteins and metabolites from cells in culture.

作者信息

Sapcariu Sean C, Kanashova Tamara, Weindl Daniel, Ghelfi Jenny, Dittmar Gunnar, Hiller Karsten

机构信息

Luxembourg Centre for Systems Biomedicine, University of Luxembourg, L-4362 Esch-Belval, Luxembourg ; HICE - Helmholtz Virtual Institute of Complex Molecular Systems in Environmental Health - Aerosols and Health, Germany 1www.hice-vi.eu.

Mass Spectrometry Core Unit, Max-Delbrück Center for Molecular Medicine, 13125 Berlin, Germany ; HICE - Helmholtz Virtual Institute of Complex Molecular Systems in Environmental Health - Aerosols and Health, Germany 1www.hice-vi.eu.

出版信息

MethodsX. 2014 Jul 18;1:74-80. doi: 10.1016/j.mex.2014.07.002. eCollection 2014.

Abstract

Proper sample preparation is an integral part of all omics approaches, and can drastically impact the results of a wide number of analyses. As metabolomics and proteomics research approaches often yield complementary information, it is desirable to have a sample preparation procedure which can yield information for both types of analyses from the same cell population. This protocol explains a method for the separation and isolation of metabolites and proteins from the same biological sample, in order for downstream use in metabolomics and proteomics analyses simultaneously. In this way, two different levels of biological regulation can be studied in a single sample, minimizing the variance that would result from multiple experiments. This protocol can be used with both adherent and suspension cell cultures, and the extraction of metabolites from cellular medium is also detailed, so that cellular uptake and secretion of metabolites can be quantified. Advantages of this technique includes:1.Inexpensive and quick to perform; this method does not require any kits.2.Can be used on any cells in culture, including cell lines and primary cells extracted from living organisms.3.A wide variety of different analysis techniques can be used, adding additional value to metabolomics data analyzed from a sample; this is of high value in experimental systems biology.

摘要

恰当的样本制备是所有组学方法不可或缺的一部分,并且会极大地影响众多分析的结果。由于代谢组学和蛋白质组学研究方法常常能产生互补信息,因此需要一种样本制备程序,该程序能够从同一细胞群体中获取用于这两种分析类型的信息。本方案解释了一种从同一生物样本中分离和提取代谢物与蛋白质的方法,以便同时用于代谢组学和蛋白质组学分析的下游应用。通过这种方式,可以在单个样本中研究两种不同水平的生物调节,最大限度地减少多次实验所导致的差异。本方案可用于贴壁细胞培养和悬浮细胞培养,还详细介绍了从细胞培养基中提取代谢物的方法,从而能够对代谢物的细胞摄取和分泌进行定量。该技术的优点包括:1.成本低廉且操作快速;此方法无需任何试剂盒。2.可用于培养中的任何细胞,包括细胞系和从生物体中提取的原代细胞。3.可使用多种不同的分析技术,为从样本中分析的代谢组学数据增添额外价值;这在实验系统生物学中具有很高的价值。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bcaf/4472845/3dcca7726d86/fx1.jpg

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