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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