Author: Cao, Rui-Yuan; Xu, Yong-fen; Zhang, Tian-Hong; Yang, Jing-Jing; Yuan, Ye; Hao, Pei; Shi, Yi; Zhong, Jin; Zhong, Wu
Title: Pediatric Drug Nitazoxanide: A Potential Choice for Control of Zika Document date: 2017_2_3
ID: sbf9qi6g_15
Snippet: Vero cells were lysed on ice in Radio-Immunoprecipitation assay (RIPA) buffer with cOmplete Protease Inhibitor cocktail tablets (Roche Diagnostics GmbH, Mannheim, Germany). Proteins were mixed with 5× loading buffer, subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis, and electrotransferred onto polyvinylidene fluoride (PVDF) membranes (Millipore). The membrane was blocked with PBS-0.05% Tween 20 containing 5% skim milk and i.....
Document: Vero cells were lysed on ice in Radio-Immunoprecipitation assay (RIPA) buffer with cOmplete Protease Inhibitor cocktail tablets (Roche Diagnostics GmbH, Mannheim, Germany). Proteins were mixed with 5× loading buffer, subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis, and electrotransferred onto polyvinylidene fluoride (PVDF) membranes (Millipore). The membrane was blocked with PBS-0.05% Tween 20 containing 5% skim milk and incubated overnight at 4°C with a homemade anti-ZIKV capsid polyclonal antibody (capsid peptide sequence designed as MKNPKKKSGGFRIVC) and β-actin (Sigma, Darmstadt, Germany). The membrane was washed 3 times with PBS-Tween and subsequently incubated at room temperature for 1 hour with secondary antibodies in PBS-Tween containing 1% skim milk. The signals were analyzed by chemiluminescence using ImageQuant LAS40000mini (GE Healthcare Bio-Sciences, Pittsburgh, PA).
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