Extraction and analysis of diagnostically useful proteins from formalin-fixed, paraffin-embedded tissue sections.

Author(s): Ikeda K, Monden T, Kanoh T, Tsujie M, Izawa H, Haba A, Ohnishi T, Sekimoto M, Tomita N, Shiozaki H, Monden M

Publication: J Histochem Cytochem, 1998, Vol. 46, Page 397-403

PubMed ID: 9487122 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was optimization of a protein extraction method for FFPE tissue (colorectal cancer and colorectal adenoma).

   Summary of Findings:

   Protein extraction in RIPA buffer containing 2% SDS at 100 degrees C for 20 minutes followed by incubation at 60 degrees C for 2 hours resulted in the greatest protein yield, with an efficiency of 164.2 ug/mg dry tissue.

   Biospecimens

   • Tissue - Colorectal

   Preservative Types

   • Formalin

   Diagnoses:

   • Neoplastic - Carcinoma
   • Neoplastic - Benign

   Platform:

   Analyte	Technology Platform
   Protein	Lowry protein assay

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Analyte Extraction and Purification	Analyte purification	0.1% SDS buffer 2% SDS buffer 0 degrees C 37 degrees C 60 degrees C Preincubation at 100 degrees C for 20 min

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2. Study Purpose

   The purpose of this study was to compare total protein profiles and expression of targeted proteins among lysates extracted from FFPE and fresh frozen tissue.

   Summary of Findings:

   While protein patterns, determined by SDS-PAGE, differed among frozen and FFPE specimen lysates, a higher degree of similarity was observed among proteins in the 10 to 120 kDa size range. Western blot analysis for eight antigens, including nuclear, cytosolic, and membrane-bound proteins ranging in size from 33 to 120 kDa produced equivalent results among protein lysates from frozen and FFPE specimens.

   Biospecimens

   • Tissue - Colorectal

   Preservative Types

   • Formalin
   • Frozen

   Diagnoses:

   • Neoplastic - Carcinoma
   • Neoplastic - Benign

   Platform:

   Analyte	Technology Platform
   Protein	1D/2D gels
   Protein	Western blot

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Type of fixation/preservation	Formalin (buffered) Snap frozen

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