Lipid Transfection
From Bridges Lab Protocols
Revision as of 19:55, 9 March 2011 by Davebridges (Talk | contribs) (Created page with '==Materials== * Shuttle Kit from Echelon [http://www.echelon-inc.com/commerce/ccc1021-shuttle-pip-kits.htm] * I used C8 lipids and dissolved to 1 mM in PBS and frozen in small al...')
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
