Selected article for: "lysate protein and lysis buffer"
Author: Hernandez, Nicholas; Melki, Isabelle; Jing, Huie; Habib, Tanwir; Huang, Susie S.Y.; Danielson, Jeffrey; Kula, Tomasz; Drutman, Scott; Belkaya, Serkan; Rattina, Vimel; Lorenzo-Diaz, Lazaro; Boulai, Anais; Rose, Yoann; Kitabayashi, Naoki; Rodero, Mathieu P.; Dumaine, Cecile; Blanche, Stéphane; Lebras, Marie-Noëlle; Leung, Man Chun; Mathew, Lisa Sara; Boisson, Bertrand; Zhang, Shen-Ying; Boisson-Dupuis, Stephanie; Giliani, Silvia; Chaussabel, Damien; Notarangelo, Luigi D.; Elledge, Stephen J.; Ciancanelli, Michael J.; Abel, Laurent; Zhang, Qian; Marr, Nico; Crow, Yanick J.; Su, Helen C.; Casanova, Jean-Laurent
Title: Life-threatening influenza pneumonitis in a child with inherited IRF9 deficiency
  • Document date: 2018_10_1
    • ID: jqv0lyfx_39
    Snippet: Fibroblasts were pretreated with IFN-α2b (Schering) or IFN-γ (Imukin; Boehringer Ingelheim) for the specified times before lysis. Cells were lysed in NP-40 lysis buffer (280 mM NaCl, 50 mM Tris, pH 8, 0.2 mM EDTA, 2 mM EGTA, 10% glycerol, 0.5% NP-40) supplemented with 1 mM dithiothreitol, PhosSTOP (Roche), and Complete protease inhibitor cocktail (Roche). 20 μg protein lysate per lane was resolved by SDS-PAGE and transferred to polyvinylidene .....
    Document: Fibroblasts were pretreated with IFN-α2b (Schering) or IFN-γ (Imukin; Boehringer Ingelheim) for the specified times before lysis. Cells were lysed in NP-40 lysis buffer (280 mM NaCl, 50 mM Tris, pH 8, 0.2 mM EDTA, 2 mM EGTA, 10% glycerol, 0.5% NP-40) supplemented with 1 mM dithiothreitol, PhosSTOP (Roche), and Complete protease inhibitor cocktail (Roche). 20 μg protein lysate per lane was resolved by SDS-PAGE and transferred to polyvinylidene fluoride membrane, which was probed with unconjugated primary antibodies and HRP-conjugated secondary antibodies. Anti-GAP DH (Santa Cruz Biotechnology) or anti-Lamin A/C (Santa Cruz Biotechnology) antibodies were used as loading controls. IRF9 was probed with antibody recognizing the C-terminus at a dilution of 1:1,000 (ISGF-3γ p48 [H10]; Santa Cruz Biotechnology). SuperSignal West Pico Chemiluminescent substrate (Thermo Fisher Scientific) was used to visualize HRP activity, and this signal was detected by an Amersham Imager 600 (GE Life Sciences).

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