Simultaneous extraction of proteins and metabolites from cells in culture
| Autor: | Daniel Weindl, Sean C. Sapcariu, Jenny Ghelfi, Karsten Hiller, Gunnar Dittmar, Tamara Kanashova |
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| Přispěvatelé: | Luxembourg Centre for Systems Biomedicine (LCSB): Metabolomics (Hiller Group) [research center] |
| Rok vydání: | 2014 |
| Předmět: |
Proteomics
Systems biology Clinical Biochemistry Population Sample preparation Computational biology Biochemistry biophysics & molecular biology [F05] [Life sciences] Biology computer.software_genre Article Isolation Metabolomics Biomolecule extraction Liquid chromatography–mass spectrometry lcsh:Science Biochimie biophysique & biologie moléculaire [F05] [Sciences du vivant] education Protocol (science) education.field_of_study GC/MS fungi food and beverages LC/MS Medical Laboratory Technology lcsh:Q Data mining Technology Platforms Biological regulation computer |
| Zdroj: | MethodsX. Elsevier (2014). MethodsX MethodsX, Vol 1, Iss C, Pp 74-80 (2014) |
| ISSN: | 2215-0161 |
| DOI: | 10.1016/j.mex.2014.07.002 |
| Popis: | Graphical abstract Three-phase methanol–water–chloroform extraction for biological samples. Examples of components available from each phase are shown. These different phases can be then used for a variety of different analysis methods on different levels of cellular regulation. 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
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