Comparison of protein precipitation methods for sample preparation prior to proteomic analysis
Protein samples should be free of salt and other disturbing agents and have an appropriate concentration to be suitable for two-dimensional (2D) electrophoresis, the principal step of proteomics. To find the most efficient method for sample preparation, we used human plasma and compared four widely...
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| Veröffentlicht in: | Journal of Chromatography A 2004-01, Vol.1023 (2), p.317-320 |
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| container_title | Journal of Chromatography A |
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| creator | Jiang, Lei He, Lin Fountoulakis, Michael |
| description | Protein samples should be free of salt and other disturbing agents and have an appropriate concentration to be suitable for two-dimensional (2D) electrophoresis, the principal step of proteomics. To find the most efficient method for sample preparation, we used human plasma and compared four widely applied precipitation methods, using trichloroacetic acid (TCA), acetone, chloroform/methanol and ammonium sulfate, as well as ultrafiltration. Precipitation with TCA and acetone and ultrafiltration resulted in an efficient sample concentration and desalting. We also found that ammonium sulfate fractionation can efficiently remove albumin, which represents more than 50% of plasma proteins. |
| doi_str_mv | 10.1016/j.chroma.2003.10.029 |
| format | Article |
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| language | eng |
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| subjects | Ammonium Sulfate - chemistry Analytical, structural and metabolic biochemistry Biological and medical sciences Blood Proteins - chemistry Blood Proteins - isolation & purification Chemical Precipitation Electrophoresis, Gel, Two-Dimensional Fundamental and applied biological sciences. Psychology General aspects, investigation methods Proteins Proteome Proteomics Sample preparation |
| title | Comparison of protein precipitation methods for sample preparation prior to proteomic analysis |
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