Selected article for: "1x Protease inhibitor cocktail and inhibitor cocktail"

Author: Sneha Rath; Eliza Prangley; Jesse Donovan; Kaitlin Demarest; Yigal Meir; Ned Wingreen; Alexei Korennykh
Title: Concerted 2-5A-Mediated mRNA Decay and Transcription Reprogram Protein Synthesis in dsRNA Response
  • Document date: 2018_12_4
  • ID: ng5c7xai_51
    Snippet: Magnetic protein A beads were incubated with 2 g anti-FLAG (Sigma) or anti-eIF2α (Santa Cruz) antibodies in IP buffer (10 mM HEPES (pH 7.5), 150 mM NaCl, 0.1% Nonidet P-40 (NP-40), 1x complete protease inhibitor cocktail, RNase inhibitor) at 4°C for two hours. Excess unbound antibody was removed by washing the beads twice in IP buffer for 5 minutes. Cells transfected with or without 2-5A for three hours were lysed in 10 mM HEPES (pH 7.5), 10.....
    Document: Magnetic protein A beads were incubated with 2 g anti-FLAG (Sigma) or anti-eIF2α (Santa Cruz) antibodies in IP buffer (10 mM HEPES (pH 7.5), 150 mM NaCl, 0.1% Nonidet P-40 (NP-40), 1x complete protease inhibitor cocktail, RNase inhibitor) at 4°C for two hours. Excess unbound antibody was removed by washing the beads twice in IP buffer for 5 minutes. Cells transfected with or without 2-5A for three hours were lysed in 10 mM HEPES (pH 7.5), 10 mM NaCl, 2 mM EDTA, 0.5% Triton X-100, 1x complete protease inhibitor cocktail and RNase inhibitor for 7 minutes while rotating, at 4 °C.

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