The impact of ultraviolet- and infrared-based laser microdissection technology on phosphoprotein detection in the laser microdissection-reverse phase protein array workflow
Abstract
Reversible protein phosphorylation represents a key mechanism by which signals are transduced in eukaryotic cells. Dysregulated phosphorylation is also a hallmark of carcinogenesis and represents key drug targets in the precision medicine space. Thus, methods that preserve phosphoprotein integrity in the context of clinical tissue analyses are crucially important in cancer research. Here we investigated the impact of UV laser microdissection (UV LMD) and IR laser capture microdissection (IR LCM) on phosphoprotein abundance of key cancer signaling protein targets assessed by reverse-phase protein microarray (RPPA). Tumor epithelial cells from consecutive thin sections obtained from four high-grade serous ovarian cancers were harvested using either UV LMD or IR LCM methods. Phosphoprotein abundances for ten phosphoproteins that represent important drug targets were assessed by RPPA and revealed no significant differences in phosphoprotein integrity from those obtained using higher-energy UV versus the lower-energy IR laser methods.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Mar 09, 2020
- Source ID
- 10.1186/s12014-020-09272-z
Entities
People
- Allison L. Hunt
- Dave Mitchell
- Elisa Baldelli
- Emanuel F. Petricoin
- G. Larry Maxwell
- Glenn Gist
- Julie Oliver
- Mariaelena Pierobon
- Nicholas W Bateman
- Thomas P Conrads
Organizations
- Uniformed Services University of the Health Sciences
- United States Department of Defense