Author: Munday, Diane C.; Emmott, Edward; Surtees, Rebecca; Lardeau, Charles-Hugues; Wu, Weining; Duprex, W. Paul; Dove, Brian K.; Barr, John N.; Hiscox, Julian A.
Title: Quantitative Proteomic Analysis of A549 Cells Infected with Human Respiratory Syncytial Virus Document date: 2010_7_20
ID: 2zhaknbi_9
Snippet: Gel Electrophoresis and In-gel Digestion-Each sample was reduced in SDS-PAGE loading buffer containing 10 mM DTT and alkylated in 50 mM iodoacetamide prior to being boiled, then separated by one-dimensional SDS-PAGE (4 -12% Bis-Tris Novex minigel, Invitrogen), and visualized by colloidal Coomassie staining (Novex, Invitrogen). The entire protein gel lane was excised and cut into 10 gel slices each. Gel slices were subjected to in-gel digestion wi.....
Document: Gel Electrophoresis and In-gel Digestion-Each sample was reduced in SDS-PAGE loading buffer containing 10 mM DTT and alkylated in 50 mM iodoacetamide prior to being boiled, then separated by one-dimensional SDS-PAGE (4 -12% Bis-Tris Novex minigel, Invitrogen), and visualized by colloidal Coomassie staining (Novex, Invitrogen). The entire protein gel lane was excised and cut into 10 gel slices each. Gel slices were subjected to in-gel digestion with trypsin (36) . The resulting tryptic peptides were extracted by 1% formic acid, acetonitrile; lyophilized in a SpeedVac (Helena Biosciences); and resuspended in 1% formic acid.
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