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Updated: May 4, 2026
Determination of Protein Expression Level in Cultured Cells by Immunocytochemistry on Paraffin-embedded Cell Blocks
Published on: May 20, 2018
Protein extraction from methanol fixed paraffin embedded tissue blocks: A new possibility using cell blocks
Theresa J Kokkat1, Diane McGarvey1, Miral S Patel1
1Address: Department of Pathology and Lab Medicine, Cooperative Human Tissue Network-Eastern Division, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
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.
