Advancing Epigenetics Towards Systems Biology

Complex Purification / Proteomics

Zuzana Jasencakova and Anja Groth

Introduction

Here we describe a strategy for isolation of multiprotein complexes from human HeLa S3 cells in a scale and purity optimized for characterization by mass spectrometry. For this purpose, we use stably expressed One-STrEP-tag ® fusion proteins. This approach was successfully used in characterization of histone chaperone Asf1 complexes (Groth et al., 2007), and we have recently optimized it further. Using this protocol we routinely obtain complexes in amounts sufficient for visualising single protein bands by Coomassie Blue staining. [...]

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Zuzana Jasencakova and Anja Groth

BRIC (Biotech Research and Innovation Centre) - University of Copenhagen - Ole Maaløes Vej 5 - DK-2200 Copenhagen N, Denmark

Zuzana Jasencakova and Anja Groth