Selected article for: "HPLC analysis and isocratic elution"
Author: Karen N. Barnard; Brynn K. Alford-Lawrence; David W. Buchholz; Brian R. Wasik; Justin R. LaClair; Hai Yu; Rebekah Honce; Stefan Ruhl; Petar Pajic; Erin K. Daugherity; Xi Chen; Stacey L. Schultz-Cherry; Hector C. Aguilar; Ajit Varki; Colin R. Parrish
Title: The effects of modified sialic acids on mucus and erythrocytes on influenza A virus HA and NA functions
  • Document date: 2019_10_10
    • ID: mgdqixt0_39
    Snippet: The sialic acid composition from tissues, mucin, and erythrocytes were analyzed by HPLC analysis as previously described (45, 76) . In brief, Sia from 20-30 mg of tissue, 50 µg of mucin or saliva, or 100 µl of washed 5% v/v erythrocytes were 475 release using 2M acetic acid at 80°C for 3 hr followed by filtration through a 10kD centrifugal filter (Microcon) and dried using a vacuum concentrator (SpeedVac). Released Sia were labeled with 1,2-di.....
    Document: The sialic acid composition from tissues, mucin, and erythrocytes were analyzed by HPLC analysis as previously described (45, 76) . In brief, Sia from 20-30 mg of tissue, 50 µg of mucin or saliva, or 100 µl of washed 5% v/v erythrocytes were 475 release using 2M acetic acid at 80°C for 3 hr followed by filtration through a 10kD centrifugal filter (Microcon) and dried using a vacuum concentrator (SpeedVac). Released Sia were labeled with 1,2-diamino-4, 5-methylenedioxybenzene (DMB, Sigma Aldrich) for 2.5 hr at 50°C. HPLC analysis was performed using a Dionex UltiMate 3000 system with an Acclaim C18 column (ThermoFisher) under isocratic elution in 7% methanol, 7% acetonitrile, and 86% water. Sia ManNAc6NAc (62) as the donor precursor using a one-pot multienzyme sialylation system similar to that described previously (79) .

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