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
Cytojournaljournal_title
CytoJournalauthors
Kokkat TJ,McGarvey D,Patel MS,Tieniber AD,Livolsi VA,Baloch ZWdoi
10.4103/1742-6413.122299subject
Has Abstractpub_date
2013-11-28 00:00:00pages
23eissn
0974-5963issn
1742-6413pii
CJ-10-23journal_volume
10pub_type
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