Journal article

A robust method for proteomic characterization of mouse cartilage using solubility-based sequential fractionation and two-dimensional gel electrophoresis

Richard Wilson, John F Bateman

MATRIX BIOLOGY | ELSEVIER SCIENCE BV | Published : 2008

Abstract

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 glob..

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Grants

Funding Acknowledgements

We are grateful to Profs. Bruce Caterson, Mats Paulsson, Dick Heinegard and the late Rupert Timpl for the provision of the antibodies used in this Study.This work was supported by funding from the NH&MRC-EU Collaborative Grant Scheme, Arthritis Australia and the Bone Growth Foundation.