Surface Plasmon Resonance - Protein Lipid Interactions: Difference between revisions
Updated protocol with more detailed notes. |
updated protocol to make 300 uL of lipid to make extruding easier |
||
| Line 15: | Line 15: | ||
==Preparation of Liposomes== | ==Preparation of Liposomes== | ||
#Combine DOPC + C16 lipid (or DOPC alone) at a ratio of 97:3 and dry under nitrogen ( | #Combine DOPC + C16 lipid (or DOPC alone) at a ratio of 97:3 and dry under nitrogen (30 uL PC or 29.1 uL PC + 9 uL PI) | ||
#Resuspend to 10 mM total lipid with HBS-N ( | #Resuspend to 10 mM total lipid with HBS-N (300 uL). Vortex thoroughly and sonicate in water bath | ||
#Correct pH to 7.4 using pH paper and | #Correct pH to 7.4 using pH paper and (usually ~ 1uL of 1M NaOH) | ||
#Freeze thaw in a water bath sonicator at 40C and liquid nitrogen 8 times. | #Freeze thaw in a water bath sonicator at 40C and liquid nitrogen 8 times. | ||
#Pass 10X through a polycarbonate filter using an Avanti MiniExtruder at Room Temperature. | #Pass 10X through a polycarbonate filter using an Avanti MiniExtruder at Room Temperature. | ||