Lipid Transfection

Materials

  • Shuttle Kit from Echelon [1]
  • I used C8 lipids and dissolved to 1 mM in PBS and frozen in small aliquots at -80.
  • Dissolved the appropriate carrier to 1 mM in PBS and freeze at -80 in small aliquots
  • For PIP use Carrier 3, for PIP2 use Carrier 2, for PIP3 use Carrier 2

Procedure

  • serum starved my cells for 3h in DMEM + PSG, using a small volume of media (~200 uL in a 12 well dish)
  • combined lipid and carrier 1:1 making enough to bring the final concentration on the cells to 1 uM
  • incubated lipid and carrier on the bench for 30 min
  • added mixture to cells for 30 min