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Life Sciences Commons

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Horticulture

University of Kentucky

Horticulture Faculty Publications

2015

Arabidopsis

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[14C] Glucose Cell Wall Incorporation Assay For The Estimation Of Cellulose Biosynthesis, Chad Brabham, Jozsef Stork, Seth Debolt Sep 2015

[14C] Glucose Cell Wall Incorporation Assay For The Estimation Of Cellulose Biosynthesis, Chad Brabham, Jozsef Stork, Seth Debolt

Horticulture Faculty Publications

Cellulose is synthesized by Cellulose Synthase A proteins at the plasma membrane using the substrate UDP glucose. Herein, we provide a detailed method for measuring the incorporation of radiolabeled glucose into the cellulose fraction of the cell wall. In this method Arabidopsis seedlings are treated for 2 h with a cellulose biosynthesis inhibitor in the presence of radiolabeled glucose, and are subsequently boiled in acetic-nitric acid to solubilize non-cellulosic material. The radiolabeled glucose detected in the insoluble fraction indicates the amount of cellulose synthesized during the experimental timeframe. The short-term nature of this method is a useful tool in determining …