Selected article for: "bovine serum and ph hepes"
Author: Takenouchi, Takato; Kitani, Hiroshi; Suzuki, Shunichi; Nakai, Michiko; Fuchimoto, Dai-ichiro; Tsukimoto, Mitsutoshi; Shinkai, Hiroki; Sato, Mitsuru; Uenishi, Hirohide
Title: Immortalization and Characterization of Porcine Macrophages That Had Been Transduced with Lentiviral Vectors Encoding the SV40 Large T Antigen and Porcine Telomerase Reverse Transcriptase
  • Document date: 2017_8_21
    • ID: 0cqtxqjv_27
    Snippet: The macrophages (3 × 10 5 cells/well in a 24-well plate) were primed with 1 µg/ml LPS for 4 h, and then the medium was replaced with 250 µl HEPES-buffered salt solution (145 mM NaCl, 2.5 mM KCl, 1 mM MgCl2, 1.8 mM CaCl2, 20 mM HEPES, 10 mM glucose, and 0.01% bovine serum albumin; pH 7.4) containing nigericin (Sigma) or ATP (Sigma). After being incubated at 37°C for 30 min, the supernatants were collected, and the cells were lysed with 200 µl.....
    Document: The macrophages (3 × 10 5 cells/well in a 24-well plate) were primed with 1 µg/ml LPS for 4 h, and then the medium was replaced with 250 µl HEPES-buffered salt solution (145 mM NaCl, 2.5 mM KCl, 1 mM MgCl2, 1.8 mM CaCl2, 20 mM HEPES, 10 mM glucose, and 0.01% bovine serum albumin; pH 7.4) containing nigericin (Sigma) or ATP (Sigma). After being incubated at 37°C for 30 min, the supernatants were collected, and the cells were lysed with 200 µl ice-cold lysis buffer. Maturation and secretion of interleukin (IL)-1β were evaluated using immunoblotting, as described previously (17) . The target protein was revealed using ImmunoStar LD (Wako) and detected using a C-DiGit blot scanner (LI-COR, Inc., Lincoln, NE, USA).

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