Pre-extraction sample handling by automated frozen disruption significantly improves subsequent proteomic analyses |
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Authors: | Butt R Hussain Coorssen Jens R |
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Affiliation: | Department of Physiology & Biophysics, Faculty of Medicine, University of Calgary, Calgary, AB, Canada. |
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Abstract: | 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. |
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