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Quantitative Proteomic Analysis of Mass Limited Tissue Samples for Spatially Resolved Tissue Profiling
Traditionally, proteomic studies have been carried out on whole tissues or organs enabling the profiling of thousands of proteins within a single LC-MS analysis. A disadvantage of this approach is that proteomes generated from whole tissues are an “average” that represents a blend of cell types and distinct anatomical regions which can obscure important biological phenomena. Laser capture microdissection (LCM) is an elegant method that allows tissue features of interest, as small as a single cell, to be identified and isolated for downstream analysis. Herein we describe an approach that utilizes an immobilized enzyme reactor (IMER) coupled directly to nanoLC-MS/MS for highly sensitive, automated, quantitative proteomic analysis of the microscopic tissue specimens generated by LCM.
KeywordsImmobilized enzyme reactor Laser capture microdissection Mass spectrometry NanoLC Nanoproteomics Proteomics
Portions of this research were supported by grants from the National Heart Lung Blood Institute of NIH (U01 HL122703), National Institute of General Medical Sciences of NIH (P41 GM103493) and the PNNL Laboratory-Directed Research and Development (LDRD) program. Work was performed in W. R. Wiley Environmental Molecular Sciences Laboratory (EMSL), a Department Of Energy (DOE) office of Biological and Environmental Research (BER) national user facility located at Pacific Northwest National Laboratory (PNNL).
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