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  29. After 1824 h, the plates were harvested on a FilterMate harvester and analyzed on a 1450 LSC Microbeta TriLux counter
  30. After 24 h in culture, supernatants have been removed and placed on microtiter plates coated with purified anti-IL-2 overnight at 4uC
  31. After 24 h incubation, in HepG2 cells, antiproliferation EC50 for Where Scontrol is the percent of supercoiled DNA in the control lane
  32. After 4 washes, antibody visualization was achieved by the incubation for 3 h at room temperature with Alexa 488-conjugated anti-mouse and/or Alexa 568-conjugated anti-rabbit IgGs
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  48. After approximately 16 hours of incubation with hormone, cells in each well were lysed using 100L mammalian protein extraction reagent
  49. After blocking, sections were stained with antibodies or phalloidin and imaged with a Leica DM4000B fluorescent microscope
  50. After centrifugation, the halo containing PBMCs was carefully transferred into a clean centrifuge tube and washed twice with 7 ml of PBS

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