Optimized Protocol for Protein Extraction from the Breast Tissue that is Compatible with Two-Dimensional Gel
Olena Zakharchenko1, Christina Greenwood, Louise Alldridge
1Karolinska Institutet, Dept. Oncology-Pathology, Karolinska Biomics Center, Cancer Translational Research Unit, Stockholm, Sweden.
Abstract:
Proteomics is a highly informative approach to analyze cancer-associated transformation in tissues. The main challenge to use a tissue for proteomics studies is the small sample size and difficulties to extract and preserve proteins. The choice of a buffer compatible with proteomics applications is also a challenge. Here we describe a protocol optimized for the most efficient extraction of proteins from the human breast tissue in a buffer compatible with two-dimensional gel electrophoresis (2D-GE). This protocol is based on mechanically assisted disintegration of tissues directly in the 2D-GE buffer. Our method is simple, robust and easy to apply in clinical practice. We demonstrate high quality of separation of proteins prepared according to the reported here protocol.
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