proteomics;
membrane proteins;
tissue homogenization;
protein extraction;
D O I:
10.1021/pr0503634
中图分类号:
Q5 [生物化学];
学科分类号:
071010 ;
081704 ;
摘要:
Here we quantitatively characterize two common homogenization strategies in the analysis of tissue proteomes: classical manual homogenization (MH) and an automated frozen disruption (AFD) technique. In a variety of tissues, many proteins were more efficiently extracted, resolved and detected, with high reproducibility after AFD, amounting to as much as 2% of the total resolved proteome. The benefits of AFD over MH are 2-fold: (1) AFD yields a much more thorough homogenate than MH; and (2) as a deep frozen alternative, AFD maintains a level of biological complexity that is not retained during MH. Thus, AFD coupled with refined 2DE protocols and Sypro Ruby staining yields quantitative proteomic analyses.
机构:
Dept. of Physiology and Biophysics, University of Calgary, Faculty of Medicine, Calgary, Alta. T2N 4N1Dept. of Physiology and Biophysics, University of Calgary, Faculty of Medicine, Calgary, Alta. T2N 4N1
Churchward M.A.
Butt R.H.
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机构:
Dept. of Physiology and Biophysics, University of Calgary, Faculty of Medicine, Calgary, Alta. T2N 4N1Dept. of Physiology and Biophysics, University of Calgary, Faculty of Medicine, Calgary, Alta. T2N 4N1
Butt R.H.
Lang J.C.
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机构:
Dept. of Physiology and Biophysics, University of Calgary, Faculty of Medicine, Calgary, Alta. T2N 4N1Dept. of Physiology and Biophysics, University of Calgary, Faculty of Medicine, Calgary, Alta. T2N 4N1
Lang J.C.
Hsu K.K.
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h-index: 0
机构:
Dept. of Physiology and Biophysics, University of Calgary, Faculty of Medicine, Calgary, Alta. T2N 4N1Dept. of Physiology and Biophysics, University of Calgary, Faculty of Medicine, Calgary, Alta. T2N 4N1
Hsu K.K.
Coorssen J.R.
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h-index: 0
机构:
Dept. of Physiology and Biophysics, University of Calgary, Faculty of Medicine, Calgary, Alta. T2N 4N1
Dept. of Biochemistry and Molecular Biology, University of Calgary, Faculty of Medicine, Calgary, Alta. T2N 4N1
Hotchkiss Brain Institute, University of Calgary, Faculty of Medicine, Calgary, Alta. T2N 4N1Dept. of Physiology and Biophysics, University of Calgary, Faculty of Medicine, Calgary, Alta. T2N 4N1
机构:
Dept. of Physiology and Biophysics, University of Calgary, Faculty of Medicine, Calgary, Alta. T2N 4N1Dept. of Physiology and Biophysics, University of Calgary, Faculty of Medicine, Calgary, Alta. T2N 4N1
Churchward M.A.
Butt R.H.
论文数: 0引用数: 0
h-index: 0
机构:
Dept. of Physiology and Biophysics, University of Calgary, Faculty of Medicine, Calgary, Alta. T2N 4N1Dept. of Physiology and Biophysics, University of Calgary, Faculty of Medicine, Calgary, Alta. T2N 4N1
Butt R.H.
Lang J.C.
论文数: 0引用数: 0
h-index: 0
机构:
Dept. of Physiology and Biophysics, University of Calgary, Faculty of Medicine, Calgary, Alta. T2N 4N1Dept. of Physiology and Biophysics, University of Calgary, Faculty of Medicine, Calgary, Alta. T2N 4N1
Lang J.C.
Hsu K.K.
论文数: 0引用数: 0
h-index: 0
机构:
Dept. of Physiology and Biophysics, University of Calgary, Faculty of Medicine, Calgary, Alta. T2N 4N1Dept. of Physiology and Biophysics, University of Calgary, Faculty of Medicine, Calgary, Alta. T2N 4N1
Hsu K.K.
Coorssen J.R.
论文数: 0引用数: 0
h-index: 0
机构:
Dept. of Physiology and Biophysics, University of Calgary, Faculty of Medicine, Calgary, Alta. T2N 4N1
Dept. of Biochemistry and Molecular Biology, University of Calgary, Faculty of Medicine, Calgary, Alta. T2N 4N1
Hotchkiss Brain Institute, University of Calgary, Faculty of Medicine, Calgary, Alta. T2N 4N1Dept. of Physiology and Biophysics, University of Calgary, Faculty of Medicine, Calgary, Alta. T2N 4N1