Improved protein extraction and protein identification from archival formalin-fixed paraffin-embedded human aortas.

ABSTRACT:

Purpose

Evaluate combination of heat and elevated pressure to enhance protein extraction and quality of formalin-fixed (FF), and FF paraffin-embedded (FFPE) aorta for proteomics.

Experiment design

Proteins were extracted from fresh frozen aorta at room temperature (RT). FF and FFPE aortas (3 months and 15 years) were extracted at RT, heat alone, or a combination of heat and high pressure. Protein yields were compared, and digested peptides from the extracts were analyzed with MS.

Results

Combined heat and elevated pressure increased protein yield from human FF or FFPE aorta compared to matched tissues with heat alone (1.5-fold) or at RT (8.3-fold), resulting in more proteins identified and with more sequence coverage. The length of storage did adversely affect the qua

SUBMITTER: Fu Z 

PROVIDER: S-EPMC4340701 | biostudies-literature | 2013 Apr

REPOSITORIES: biostudies-literature