A rapid extraction method for glycogen from formalin-fixed liver

Sullivan, Mitchell A., Li, Shihan, Aroney, Samuel T.N., Deng, Bin, Li, Cheng, Roura, Eugeni, Schulz, Benjamin L., Harcourt, Brooke E., Forbes, Josephine M. and Gilbert, Robert G. (2015) A rapid extraction method for glycogen from formalin-fixed liver. Carbohydrate Polymers, 118 9-15. doi:10.1016/j.carbpol.2014.11.005

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Author Sullivan, Mitchell A.
Li, Shihan
Aroney, Samuel T.N.
Deng, Bin
Li, Cheng
Roura, Eugeni
Schulz, Benjamin L.
Harcourt, Brooke E.
Forbes, Josephine M.
Gilbert, Robert G.
Title A rapid extraction method for glycogen from formalin-fixed liver
Journal name Carbohydrate Polymers   Check publisher's open access policy
ISSN 0144-8617
1879-1344
Publication date 2015-03-01
Year available 2014
Sub-type Article (original research)
DOI 10.1016/j.carbpol.2014.11.005
Open Access Status Not yet assessed
Volume 118
Start page 9
End page 15
Total pages 7
Place of publication Oxford United Kingdom
Publisher Pergamon Press
Language eng
Subject 2507 Polymers and Plastics
1605 Organic Chemistry
2505 Materials Chemistry
Abstract Liver glycogen, a highly branched polymer, acts as our blood-glucose buffer. While past structural studies have extracted glycogen from fresh or frozen tissue using a cold-water; sucrose-gradient centrifugation technique, a method for the extraction of glycogen from formalin-fixed liver would allow the analysis of glycogen from human tissues that are routinely collected in pathology laboratories. In this study, both sucrose-gradient and formalin-fixed extraction techniques were carried out on piglet livers, with the yields, purities and size distributions (using size exclusion chromatography) compared. The formalin extraction technique, when combined with a protease treatment, resulted in higher yields (but lower purities) of glycogen with size distributions similar to the sucrose-gradient centrifugation technique. This formalin extraction procedure was also significantly faster, allowing glycogen extraction throughput to increase by an order of magnitude. Both extraction techniques were compatible with mass spectrometry proteomics, with analysis showing the two techniques were highly complementary. (C) 2014 Elsevier Ltd. All rights reserved.
Formatted abstract
Liver glycogen, a highly branched polymer, acts as our blood-glucose buffer. While past structural studies have extracted glycogen from fresh or frozen tissue using a cold-water, sucrose-gradient centrifugation technique, a method for the extraction of glycogen from formalin-fixed liver would allow the analysis of glycogen from human tissues that are routinely collected in pathology laboratories. In this study, both sucrose-gradient and formalin-fixed extraction techniques were carried out on piglet livers, with the yields, purities and size distributions (using size exclusion chromatography) compared. The formalin extraction technique, when combined with a protease treatment, resulted in higher yields (but lower purities) of glycogen with size distributions similar to the sucrose-gradient centrifugation technique. This formalin extraction procedure was also significantly faster, allowing glycogen extraction throughput to increase by an order of magnitude. Both extraction techniques were compatible with mass spectrometry proteomics, with analysis showing the two techniques were highly complementary.
Keyword Formalin
Glycogen extraction
SEC
Q-Index Code C1
Q-Index Status Confirmed Code
Grant ID APP1031542
Institutional Status UQ

 
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