Pre-extraction sample handling by automated frozen disruption significantly improves subsequent proteomic analyses

被引：40

作者：

Butt, RH

Coorssen, JR

机构：

[1] Univ Calgary, Fac Med, Hotchkiss Brain Inst, Calgary, AB, Canada

[2] Univ Calgary, Fac Med, Dept Physiol & Biophys, Calgary, AB, Canada

[3] Univ Calgary, Fac Med, Dept Biochem & Mol Biol, Calgary, AB, Canada

[4] Univ Calgary, Fac Med, Dept Cell Biol & Anat, Calgary, AB, Canada

来源：

JOURNAL OF PROTEOME RESEARCH | 2006年 / 5卷 / 02期

关键词：

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.

引用

页码：437 / 448

页数：12