UA Biophysics:Protocols:Lipid Extraction

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Lipid Extration

  • Cells are harvested by centrifugation at 2800 rpm for 15 min
  • The supernatant are discard
  • Cell pellet frozen overnight at -30°C
  • Frozen biomass are lyophilize for 15h to remove the residual water
  • The samples are dissolve in a mixture of chloroform/methanol/water (3:1:1 v/v) and vortexed every 15min for 4h
  • The samples are then allowed to stand for 2 days to separate the three phases, after which the lower phase of chloroform and lipid is drawn off by aspiration and collected in a clean glass tube
  • A lipid film is formed by drying the chloroform with a gentle stream of N2 and store at -20°C until further experimentation

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