Selected article for: "activity luciferase and lysis buffer"

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_17
    Snippet: SupT1 cells were seeded at 4 X10 5 cells/ml in 12-well dishes. Infectious virus was added in the presence of 4 μg/ml polybrene with spinoculation at 1,850 X g for 1 hour at 4˚C and then incubated at 37˚C for 6 hours. SupT1 cells were then refed and incubated for an additional 48 hours. Viral supernatants were used to infect HeLa-TZM-bl cells. Forty-eight hours post-infection, HeLa-TZM-bl cells were harvested in lysis buffer (50 mM Tris pH 7.4,.....
    Document: SupT1 cells were seeded at 4 X10 5 cells/ml in 12-well dishes. Infectious virus was added in the presence of 4 μg/ml polybrene with spinoculation at 1,850 X g for 1 hour at 4˚C and then incubated at 37˚C for 6 hours. SupT1 cells were then refed and incubated for an additional 48 hours. Viral supernatants were used to infect HeLa-TZM-bl cells. Forty-eight hours post-infection, HeLa-TZM-bl cells were harvested in lysis buffer (50 mM Tris pH 7.4, 150 mM NaCl, 1 mM EDTA, 1 mM EGTA, 1% Triton X-100). HeLa-TZM-bl lysates were mixed at a 1:1 ratio with assay buffer (200 mM Tris pH 7.8, 250 mM MgCl2, 7.5 mM ATP, 25 mM CoA, 0.2 mg/ml D-Luciferin) and measured for luciferase activity using a Perkin Elmer plate reader.

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