Selected article for: "anti antibody and buffer sample"
Author: Alisha Chitrakar; Sneha Rath; Jesse Donovan; Kaitlin Demarest; Yize Li; Raghavendra Rao Sridhar; Susan R. Weiss; Sergei V. Kotenko; Ned S. Wingreen; Alexei Korennykh
Title: Realtime 2-5A kinetics suggests interferons ß and ? evade global arrest of translation by RNase L
  • Document date: 2018_11_26
    • ID: mxdvdw9u_29
    Snippet: To generate puromycin-tagged nascent peptides, human cells were treated with 0.1-1 µg/mL of poly-IC for times specified on the figures, after which the growth media was supplemented with 10 µg/mL puromycin (Invitrogen). Puromycin pulse lasted for 5 min. Cells were trypsinized and harvested in NuPAGE LDS sample buffer for western blot analyses. Proteins were separated by 10% BisTris PAGE (NuPAGE), and transferred on PVDF membranes (Life Technolo.....
    Document: To generate puromycin-tagged nascent peptides, human cells were treated with 0.1-1 µg/mL of poly-IC for times specified on the figures, after which the growth media was supplemented with 10 µg/mL puromycin (Invitrogen). Puromycin pulse lasted for 5 min. Cells were trypsinized and harvested in NuPAGE LDS sample buffer for western blot analyses. Proteins were separated by 10% BisTris PAGE (NuPAGE), and transferred on PVDF membranes (Life Technologies). The membrane was stained with Ponceau to normalize for sample loading, then washed and blocked with 5% non-fat dry milk in TBST buffer. The membranes were probed with 1:1000 mouse anti-puromycin antibody (EMD Millipore) that binds to de novo synthesized proteins, followed by horseradish peroxidase-conjugated goat anti-mouse secondary antibody (1:10,000, Jackson ImmunoResearch).

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    • µg ml and dry milk: 1, 2
    • µg ml and growth medium: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15
    • µg ml and human cell: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34
    • µg ml and poly IC ml: 1
    • µg ml and PVDF membrane: 1
    • µg ml and sample buffer: 1, 2, 3, 4, 5, 6
    • µg ml and sample loading: 1, 2
    • µg ml and secondary antibody: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10
    • µg ml and western blot: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28
    • µg ml and western blot analysis: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11
    • µg ml and western blot analysis sample buffer: 1
    • µg poly IC ml and poly IC ml: 1
    • anti mouse secondary antibody and dry milk: 1
    • anti mouse secondary antibody and Jackson ImmunoResearch anti mouse secondary antibody: 1, 2
    • anti mouse secondary antibody and PVDF membrane: 1
    • anti mouse secondary antibody and sample buffer: 1, 2
    • anti mouse secondary antibody and secondary antibody: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43
    • anti mouse secondary antibody and western blot: 1, 2, 3, 4, 5, 6
    • anti mouse secondary antibody and western blot analysis: 1, 2, 3