Author: Xing, Yang; Liqi, Zhu; Jian, Lin; Qinghua, Yu; Qian, Yang
Title: Doxycycline Induces Mitophagy and Suppresses Production of Interferon-ß in IPEC-J2 Cells Document date: 2017_2_1
ID: 1ljye9pj_19
Snippet: Cells were lysed in RIPA buffer (50 mM Tris-HCl (pH 7.4), 150 mM NaCl, 1% NP-40) containing a protease inhibitor cocktail (Yhermo Science). The protein concentration was determined. Equal amounts of protein were separated by SDS-PAGE and electrophoretically transferred onto a polyvinylidene difluoride (PVDF) membrane (Millipore, Shanghai, China). After blocking with 5% nonfat milk in Tris-buffered saline containing 0.1% Tween 20, the membrane was.....
Document: Cells were lysed in RIPA buffer (50 mM Tris-HCl (pH 7.4), 150 mM NaCl, 1% NP-40) containing a protease inhibitor cocktail (Yhermo Science). The protein concentration was determined. Equal amounts of protein were separated by SDS-PAGE and electrophoretically transferred onto a polyvinylidene difluoride (PVDF) membrane (Millipore, Shanghai, China). After blocking with 5% nonfat milk in Tris-buffered saline containing 0.1% Tween 20, the membrane was incubated with specific primary antibodies (1:1000), followed by incubation with appropriate horseradish peroxidase-conjugated secondary antibodies. Signals were detected using SuperSignal WestPico kit (Thermo Scientific) and subjected to Image Reader LAS-4000 imaging system (FUJIFLIM, Japan).
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