Selected article for: "anti mouse antibody and western blot analysis"
Author: Cong, Yingying; Kriegenburg, Franziska; de Haan, Cornelis A. M.; Reggiori, Fulvio
Title: Coronavirus nucleocapsid proteins assemble constitutively in high molecular oligomers
  • Document date: 2017_7_18
    • ID: 15hzah62_25
    Snippet: For the pull-down experiments, GSH-Sepharose bound GST fusion protein were incubated with 200 µl of bacterial extract or 200 µl of LR7 cell extracts on a rotatory wheel for 2 h at 4 °C, subsequently washed at 4 °C three times in PBS supplemented with 5 mM DTT, 10% glycerol, 1% Triton X-100 and one time in PBS buffer. Proteins bound to the Sepharose beads were eluted in 20 µl of sample buffer by boiling and subjected to SDS-PAGE, blotted onto.....
    Document: For the pull-down experiments, GSH-Sepharose bound GST fusion protein were incubated with 200 µl of bacterial extract or 200 µl of LR7 cell extracts on a rotatory wheel for 2 h at 4 °C, subsequently washed at 4 °C three times in PBS supplemented with 5 mM DTT, 10% glycerol, 1% Triton X-100 and one time in PBS buffer. Proteins bound to the Sepharose beads were eluted in 20 µl of sample buffer by boiling and subjected to SDS-PAGE, blotted onto PVDF membranes and visualized by either membrane staining with Ponceau Red or western blot analysis using anti-6xHis antibody (HIS H8, Thermo, Waltham, MA) or anti-N protein monoclonal antibodies 11 . Bound primary antibodies were detected using the Alexa680-conjugated goat polyclonal anti-mouse IgG antibody (Life Technologies) and signals visualized with an Odyssey system (LI-COR, Lincoln, NE).

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