Protein extraction from formalin-fixed, paraffin-embedded tissue sections: quality evaluation by mass (original) (raw)

Author(s): Shi SR, Liu C, Balgley BM, Lee C, Taylor CR

Publication: J Histochem Cytochem, 2006, Vol. 54, Page 739-43

PubMed ID: 16399996PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to compare protein profiles of extracts isolated from fresh frozen specimens and formalin-fixed, paraffin-embedded (FFPE) specimens subjected to antigen retrieval protocols differing in solution pH and temperature.

Conclusion of Paper

Results indicate that antigen retrieval in a solution containing 2% SDS at a pH of 7 or 9 under high heat (100 degrees C) yielded protein profiles similar to those observed with fresh frozen specimens.

  1. Study Purpose

The purpose of this study was to compare protein banding patterns, observed via SDS-PAGE analysis, of fresh frozen and FFPE specimens subjected to antigen retrieval protocols differing in solution pH and temperature.

Summary of Findings:

SDS protein profiles were comparable among freshly frozen specimens and FFPE specimens subjected to heat-induced antigen retrieval in a Tris-HCl buffer containing 2% SDS, with comparable results observed at the two solution pHs investigated. However, antigen retrieval at room temperature failed to produce observable protein bands. 2. ###### Study Purpose
The purpose of this study was to compare protein profiles obtained via capillary isoelectric focusing (CIEF) nano-reverse-phase liquid chromatography (RPLC)/ mass spectrometry among freshly frozen and FFPE specimens subjected to antigen retrieval protocols differing in solution pH and temperature.

Summary of Findings:

The majority of proteins identified were common among the differentially preserved specimens and antigen retrieval methods investigated, with 84% and 72% of proteins identified in fresh specimens also observed in FFPE specimens that underwent heat-induced retrieval in a solution of pH 7 or 9, respectively. FFPE specimens subjected to retrieval at room temperature yielded fewer identifiable proteins, representing a commonality of 54% with fresh frozen specimens. Distribution of identified proteins according to their identified subcellular locale was similar among preservation and antigen retrieval methods.

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