Author: He, Cheng; Kong, Ling; Zhou, Lijuan; Xia, Jing; Wei, Haixia; Liu, Min; Peng, Hongjuan
Title: Host Cell Vimentin Restrains Toxoplasma gondii Invasion and Phosphorylation of Vimentin is Partially Regulated by Interaction with TgROP18 Document date: 2017_9_5
ID: z9mg639h_45
Snippet: For western blotting, cells or parasites were lysed in lysis buffer (Beyotime Biotechnology) for 30 min at 4°C. Cell debris was precipitated by centrifugation at 12,000g at 4°C for 10 min. Supernatants were boiled in SDS-PAGE sample buffer and loaded for SDS-PAGE. Proteins were separated on polyacrylamide gels, and then transferred to PVDF membranes. Membranes were blocked in PBS containing 5% Bovine Serum Albumin (BSA) and 0.05% Tween-20 and p.....
Document: For western blotting, cells or parasites were lysed in lysis buffer (Beyotime Biotechnology) for 30 min at 4°C. Cell debris was precipitated by centrifugation at 12,000g at 4°C for 10 min. Supernatants were boiled in SDS-PAGE sample buffer and loaded for SDS-PAGE. Proteins were separated on polyacrylamide gels, and then transferred to PVDF membranes. Membranes were blocked in PBS containing 5% Bovine Serum Albumin (BSA) and 0.05% Tween-20 and probed with primary antibodies, followed by secondary antibodies conjugated with horseradish peroxidase (HRP) (See Supplementary Material for details). Specific proteins on membranes were visualized by luminescence generated using Clarity TM Western ECL Substrate (Bio-Rad) and photographed with a ChemiDoc TM Touch Imaging System (Bio-Rad).
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