Selected article for: "helix substrate and standard helix unwinding assay"
Author: Shu, Ting; Gan, Tianyu; Bai, Peng; Wang, Xiaotong; Qian, Qi; Zhou, Hui; Cheng, Qi; Qiu, Yang; Yin, Lei; Zhong, Jin; Zhou, Xi
Title: Ebola virus VP35 has novel NTPase and helicase-like activities
  • Document date: 2019_6_20
    • ID: u3pxycqh_17
    Snippet: The standard helix unwinding assay was performed as previously described (27) with minor modifications. Briefly, 20 pmol of recombinant protein and 0.1 pmol of HEX-labeled helix substrate were added to a mixture containing 50 mM HEPES-KOH (pH 8.0), 2 mM MgCl 2 , 100 mM NaCl 2 and 20 U RNasin (Promega). After incubation at 37 • C for 60 min, the reaction was terminated by adding 5× loading buffer [100 mM Tris-HCl, 1% SDS, 50% glycerol, and brom.....
    Document: The standard helix unwinding assay was performed as previously described (27) with minor modifications. Briefly, 20 pmol of recombinant protein and 0.1 pmol of HEX-labeled helix substrate were added to a mixture containing 50 mM HEPES-KOH (pH 8.0), 2 mM MgCl 2 , 100 mM NaCl 2 and 20 U RNasin (Promega). After incubation at 37 • C for 60 min, the reaction was terminated by adding 5× loading buffer [100 mM Tris-HCl, 1% SDS, 50% glycerol, and bromophenol blue (pH 7.5)]. The mixtures were then electrophoresed on 15% native-PAGE gels, followed by scanning with a Typhoon 9500 imager (GE Healthcare, Piscataway, NJ, USA).

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