Selected article for: "mm DTT and mm KCl"
Author: Wilkins, Jordan; Zheng, Yi-Min; Yu, Jingyou; Liang, Chen; Liu, Shan-Lu
Title: Nonhuman Primate IFITM Proteins Are Potent Inhibitors of HIV and SIV
  • Document date: 2016_6_3
    • ID: 1m1woscb_15
    Snippet: The experiments were performed as previously described [15] . Briefly, 10 μL virions containing culture supernatants were incubated with 40 μL of reaction mixture at 37°C for 3 h. The mixture contains 50 mM Tris-HCl pH 7.9, 5 mM MgCl2, 0.5 mM EGTA, 0.05% Triton X-100, 2% (V/V) ethylene glycol, 150 mM KCL, 5 mM DTT, 0.3 mM GSH (reduced glutathione), 0.5 U/ mL poly (rA) oligo (dT), and 0.1 μCi/μL 3H dTTP (Perkin-Elmer). The reaction was stoppe.....
    Document: The experiments were performed as previously described [15] . Briefly, 10 μL virions containing culture supernatants were incubated with 40 μL of reaction mixture at 37°C for 3 h. The mixture contains 50 mM Tris-HCl pH 7.9, 5 mM MgCl2, 0.5 mM EGTA, 0.05% Triton X-100, 2% (V/V) ethylene glycol, 150 mM KCL, 5 mM DTT, 0.3 mM GSH (reduced glutathione), 0.5 U/ mL poly (rA) oligo (dT), and 0.1 μCi/μL 3H dTTP (Perkin-Elmer). The reaction was stopped by adding 10% (V/V) cold Trichloroacetic Acid (TCA) at 4°C for 30 min, and the mixture was transferred to Millipore MultiScreen Glass Fiber FC plate. After being washed twice with cold 10% (V/V) TCA and cold ethanol, the membranes were inserted into Beta Gamma vials and read in Microbeta counter (Beckman Coulter).

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