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Layered electrophoretic transfer – A method for pre‐analytic processing of histological sections

Layered electrophoretic transfer – A method for pre‐analytic processing of histological sections Current technologies for measuring protein expression across a tissue section are based on MS or in situ detection such as immunohistochemistry. However, due to the inherent molecular complexity of tissue samples and the large dynamic range of protein expression in cells, current approaches are often unable to measure moderate‐ and low‐abundant proteins. In addition, they do not provide information on the physico‐chemical properties of the proteins studied. To address these problems, we are developing a new pre‐analytic methodology termed layered electrophoretic transfer (LET) that selectively separates and processes proteins from an intact tissue section without compromising important two‐dimensional histological information. LET offers two potential advantages over standard techniques: (i) A reduced complexity of the tissue proteome for subsequent analysis; (ii) An opportunity to assess the biochemical status of proteins as they exist in situ. As an initial proof‐of‐concept, we demonstrate here that the protein content from a mixture of molecular weight standards, human tissue lysates, and tissue sections can be successfully transferred and separated using LET, and further demonstrate that the method can be coupled with immunoblotting or MS for downstream measurements. LET technology represents a new pre‐analytic tool for interrogating the proteome in tissue sections while preserving valuable spatial information. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Proteomics Wiley

Layered electrophoretic transfer – A method for pre‐analytic processing of histological sections

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References (20)

Publisher
Wiley
Copyright
"Copyright © 2011 Wiley Subscription Services, Inc., A Wiley Company"
ISSN
1615-9853
eISSN
1615-9861
DOI
10.1002/pmic.201000476
pmid
21280224
Publisher site
See Article on Publisher Site

Abstract

Current technologies for measuring protein expression across a tissue section are based on MS or in situ detection such as immunohistochemistry. However, due to the inherent molecular complexity of tissue samples and the large dynamic range of protein expression in cells, current approaches are often unable to measure moderate‐ and low‐abundant proteins. In addition, they do not provide information on the physico‐chemical properties of the proteins studied. To address these problems, we are developing a new pre‐analytic methodology termed layered electrophoretic transfer (LET) that selectively separates and processes proteins from an intact tissue section without compromising important two‐dimensional histological information. LET offers two potential advantages over standard techniques: (i) A reduced complexity of the tissue proteome for subsequent analysis; (ii) An opportunity to assess the biochemical status of proteins as they exist in situ. As an initial proof‐of‐concept, we demonstrate here that the protein content from a mixture of molecular weight standards, human tissue lysates, and tissue sections can be successfully transferred and separated using LET, and further demonstrate that the method can be coupled with immunoblotting or MS for downstream measurements. LET technology represents a new pre‐analytic tool for interrogating the proteome in tissue sections while preserving valuable spatial information.

Journal

Proteomics – Wiley

Published: Mar 1, 2011

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