Selected article for: "room temperature and SDS polyacrylamide"
Author: Cao, Liyan; Ge, Xuying; Gao, Yu; Herrler, Georg; Ren, Yudong; Ren, Xiaofeng; Li, Guangxing
Title: Porcine epidemic diarrhea virus inhibits dsRNA-induced interferon-ß production in porcine intestinal epithelial cells by blockade of the RIG-I-mediated pathway
  • Document date: 2015_8_18
    • ID: 0qt7eth0_25
    Snippet: IECs were infected with PEDV at an MOI of 1 or treated with poly (I:C) for the indicative times, lysed in 2× sodium dodecyl sulfate (SDS) sample buffer and boiled for 10 min. Whole Cells extracts were separated by 12 % SDS-polyacrylamide gel electrophoresis and transferred to a nitrocellulose membrane, which was blocked with 5 % (w/v) bovine serum albumin (BSA) in tris-buffered saline (10 mM Tris-Cl at pH 7.5 and 150 mM NaCl) containing 0.05 % T.....
    Document: IECs were infected with PEDV at an MOI of 1 or treated with poly (I:C) for the indicative times, lysed in 2× sodium dodecyl sulfate (SDS) sample buffer and boiled for 10 min. Whole Cells extracts were separated by 12 % SDS-polyacrylamide gel electrophoresis and transferred to a nitrocellulose membrane, which was blocked with 5 % (w/v) bovine serum albumin (BSA) in tris-buffered saline (10 mM Tris-Cl at pH 7.5 and 150 mM NaCl) containing 0.05 % Tween 20 (TBST) at room temperature for 1 h. The membranes were then incubated with a primary antibody (dilution, 1:1000) at 4°C overnight and a secondary HRP-conjugated antibody (dilution, 1:2500) for 1 h at room temperature. Protein blots were developed using an enhanced chemiluminescence (ECL) detection system and exposed to X-ray film (Clinx Science Instruments Co., Ltd., Shanghai, China).

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