Selected article for: "lysis buffer and protein concentration"
Author: Yen, Wei-Chen; Wu, Yi-Hsuan; Wu, Chih-Ching; Lin, Hsin-Ru; Stern, Arnold; Chen, Shih-Hsiang; Shu, Jwu-Ching; Tsun-Yee Chiu, Daniel
Title: Impaired inflammasome activation and bacterial clearance in G6PD deficiency due to defective NOX/p38 MAPK/AP-1 redox signaling
  • Document date: 2019_11_2
    • ID: 6fw4thkq_22
    Snippet: The total protein in lysates and supernatants was analyzed by a Western blot. The cells were collected using lysis buffer (50 mM Tris-HCl, 150 mM NaCl 2 , 1 mM EDTA, 0.05% SDS, 1 mM NaF, 1% Triton-X 100, pH 7.5), and the protein concentration was determined by the Bradford assay. The protein in supernatants was concentrated by trichloroacetic acid (TCA) precipitation [39] . Samples were denatured, electrophoresed on SDS-polyacrylamide gel, and tr.....
    Document: The total protein in lysates and supernatants was analyzed by a Western blot. The cells were collected using lysis buffer (50 mM Tris-HCl, 150 mM NaCl 2 , 1 mM EDTA, 0.05% SDS, 1 mM NaF, 1% Triton-X 100, pH 7.5), and the protein concentration was determined by the Bradford assay. The protein in supernatants was concentrated by trichloroacetic acid (TCA) precipitation [39] . Samples were denatured, electrophoresed on SDS-polyacrylamide gel, and transferred onto PVDF membranes. The membranes were incubated overnight at 4°C with an appropriate dilution of a primary antibody (1:1000). The membranes were then incubated with an appropriate dilution of an HRP-conjugated secondary antibody for 1.5 h. The immunoreactive bands were visualized by ECL reagents. ImageJ software was used to analyze the intensity. β-Actin was used as a loading control.

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