Selected article for: "polyclonal antibody and sample buffer"
Author: Stewart, Meredith E.; Roy, Polly
Title: Structure-based identification of functional residues in the nucleoside-2'-O-methylase domain of Bluetongue virus VP4 capping enzyme
  • Document date: 2015_2_24
    • ID: vzel6r43_30
    Snippet: To determine whether the mutations in 2 0 -O MT domain affected the binding to cap0, we performed m7-GTP pull-down assay. Purified recombinant protein was incubated with m7-GTP sepharose at 4°C for at least 4 h. Samples were centrifuged at 500Âg for 5 min, unbound fraction removed and washed the beads 3 times with binding buffer (50 mM TrisHCl pH 7.5, 300 mM NaCl, 0.5% NP40) supplemented with cold GTP to remove non-specific interactions. Intera.....
    Document: To determine whether the mutations in 2 0 -O MT domain affected the binding to cap0, we performed m7-GTP pull-down assay. Purified recombinant protein was incubated with m7-GTP sepharose at 4°C for at least 4 h. Samples were centrifuged at 500Âg for 5 min, unbound fraction removed and washed the beads 3 times with binding buffer (50 mM TrisHCl pH 7.5, 300 mM NaCl, 0.5% NP40) supplemented with cold GTP to remove non-specific interactions. Interacting proteins were eluted from the beads by the addition of SDS-PAGE sample buffer. Samples were resolved and analyzed by western immunoblot using VP4 polyclonal antibody.

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