Journal
MATRIX BIOLOGY
Volume 27, Issue 8, Pages 709-712Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.matbio.2008.07.007
Keywords
Cartilage; Proteomics; 2-D electrophoresis; Sample fractionation
Categories
Funding
- NH&MRC-EU Collaborative Grant Scheme
- Arthritis Australia
- Bone Growth Foundation
Ask authors/readers for more resources
Identification of protein expression differences using two-dimensional electrophoresis (2-DE) and multidimensional liquid chromatography (MDLC)-based proteomics depends critically on reproducibility throughout sample preparation and analysis. This applies particularly where sample fractionation is used to remove high abundance or interfering components to facilitate deeper mining of the proteome. Here we present a procedure for solubility-based cartilage fractionation using sequential extraction with 1 M sodium chloride followed by 4 M guanidinium hydrochloride. We characterized the extracts by 1-D electrophoresis and immunoblotting for individual cellular and matrix components and more globally by 2-DE. In general, NaCl extracts were highly enriched for cellular proteins and GuHCl extracts were predominantly matrix components, with some interesting exceptions. Importantly, we observed high inter-sample reproducibility and strong correlation between targeted and global analysis, indicating that Our method can be applied to differential proteomic analysis of normal and pathological cartilage sub-proteomes. (C) 2008 Elsevier B.V. All rights reserved.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available