Optimized Protocol for Protein Extraction from the Breast Tissue that is Compatible with Two-Dimensional Gel Electrophoresis | Zendy

Olena Zakharchenko | Zendy; Christina Greenwood | Zendy; Louise Alldridge | Zendy; Serhiy Souchelnytskyi | Zendy

Open Access

Optimized Protocol for Protein Extraction from the Breast Tissue that is Compatible with Two-Dimensional Gel Electrophoresis

Author(s) -

Olena Zakharchenko,

Christina Greenwood,

Louise Alldridge,

Serhiy Souchelnytskyi

Publication year - 2011

Publication title -

breast cancer basic and clinical research

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.741

H-Index - 23

ISSN - 1178-2234

DOI - 10.4137/bcbcr.s6263

Subject(s) - buffer (optical fiber) , proteomics , protocol (science) , human breast , chromatography , extraction (chemistry) , computer science , protein purification , sample preparation , computational biology , chemistry , breast cancer , biology , cancer , biochemistry , medicine , pathology , telecommunications , genetics , alternative medicine , gene

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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