Author: Barnard, Karen N.; Wasik, Brian R.; LaClair, Justin R.; Buchholz, David W.; Weichert, Wendy S.; Alford-Lawrence, Brynn K.; Aguilar, Hector C.; Parrish, Colin R.
Title: Expression of 9-O- and 7,9-O-Acetyl Modified Sialic Acid in Cells and Their Effects on Influenza Viruses Document date: 2019_12_3
ID: 11ejfiwe_30
Snippet: Quantification of Sia variants. The Sia composition of cells were determined by incubating with 2 M acetic acid at 80°C for 3 h, filtration through a Microcon 10-kDa centrifugal filter (Millipore), and drying in a SpeedVac vacuum concentrator. Released Sia were derivatized with DMB (Sigma-Aldrich) for 2.5 h at 50°C (39) . HPLC analysis was performed using a Dionex UltiMate 3000 system with an Acclaim C 18 column (Thermo Fisher) under isocratic .....
Document: Quantification of Sia variants. The Sia composition of cells were determined by incubating with 2 M acetic acid at 80°C for 3 h, filtration through a Microcon 10-kDa centrifugal filter (Millipore), and drying in a SpeedVac vacuum concentrator. Released Sia were derivatized with DMB (Sigma-Aldrich) for 2.5 h at 50°C (39) . HPLC analysis was performed using a Dionex UltiMate 3000 system with an Acclaim C 18 column (Thermo Fisher) under isocratic elution in 7% methanol, 7% acetonitrile, and 86% water. Sia standards included bovine submaxillary mucin and commercial standards for Neu5Ac and Neu5Gc (Sigma-Aldrich). Statistical analyses were performed in GraphPad Prism software (v8).
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