Protein extraction from methanol fixed paraffin embedded tissue blocks: A new possibility using cell blocks.

Abstract:

BACKGROUND:Methanol fixed and paraffin embedded (MFPE) cellblocks are an essential cytology preparation. However, MFPE cellblocks often contain limited material and their relatively small size has caused them to be overlooked in biomarker discovery. Advances in the field of molecular biotechnology have made it possible to extract proteins from formalin fixed and paraffin embedded (FFPE) tissue blocks. In contrast, there are no established methods for extracting proteins from MFPE cellblocks. We investigated commonly available CHAPS (3-[(3-cholamidopropyl) dimethylammonio]-1-propanesulfonate) buffer, as well as two commercially available Qiagen(®) kits and compared their effectiveness on MFPE tissue for protein yields. MATERIALS AND METHODS:MFPE blocks were made by Cellient™ automated system using human tissue specimens from normal and malignant specimens collected in ThinPrep™ Vials. Protein was extracted from Cellient-methanol fixed and paraffin embedded blocks with CHAPS buffer method as well as FFPE and Mammalian Qiagen(®) kits. RESULTS:Comparison of protein yields demonstrated the effectiveness of various protein extraction methods on MFPE cellblocks. CONCLUSION:In the current era of minimally invasive techniques to obtain minimal amount of tissue for diagnostic and prognostic purposes, the use of commercial and lab made buffer on low weight MFPE scrapings obtained by Cellient(®) processor opens new possibilities for protein biomarker research.

journal_name

Cytojournal

journal_title

CytoJournal

authors

Kokkat TJ,McGarvey D,Patel MS,Tieniber AD,Livolsi VA,Baloch ZW

doi

10.4103/1742-6413.122299

subject

Has Abstract

pub_date

2013-11-28 00:00:00

pages

23

eissn

0974-5963

issn

1742-6413

pii

CJ-10-23

journal_volume

10

pub_type

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