In vitro assays are invaluable for the biochemical characterization of UDP-sugar:undecaprenyl-phosphate sugar-1-phosphate transferases. These assays typically involve the use of a radiolabeled substrate and subsequent extraction of the product, which resides in a lipid environment. Here, we describe the preparation of bacterial membranes containing these enzymes, a standard in vitro transferase assay with solvents containing chloroform and methanol, and two methods to measure product formation: scintillation counting and thin layer chromatography. © Springer Science+Business Media New York 2013.
CITATION STYLE
Patel, K. B., & Valvano, M. A. (2013). In vitro UDP-Sugar: Undecaprenyl-phosphate sugar-1-phosphate transferase assay and product detection by thin layer chromatography. Methods in Molecular Biology, 1022, 173–183. https://doi.org/10.1007/978-1-62703-465-4_14
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