Admin: Created page with "= Protocol = ===== Lipid Preparation: ===== #In a glass container, and using lipids dissolved in chloroform, mix desired lipids to a final quantity of ~4mmol total lipid. #..."

2021-02-01T19:58:23Z

Created page with "= Protocol = ===== Lipid Preparation: ===== #In a glass container, and using lipids dissolved in chloroform, mix desired lipids to a final quantity of ~4mmol total lipid. #..."

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= Protocol =

===== Lipid Preparation: =====

#In a glass container, and using lipids dissolved in chloroform, mix desired lipids to a final quantity of ~4mmol total lipid.
#Dry lipids under inert gas.
#Suspend lipids in 400ml PBS or other buffer, sonicate to suspend completely.

*Note: if covered with inert gas, these lipids are stable for ~1 week at 4oC. 

===== Glass Bead Preparation: =====

#Measure out 2mg of glass beads of the desired diameter.
#Wash 3X in 1ml ddH2O.
#Pellet glass beads. Remove as much water as possible.
#Suspend beads in 400ml of the PBS-lipid solution, rotate at room temp 20min to allow bylayers to self-assemble.
#Pellet beads with a brief (<5sec) cetrifugation and remove the liquid phase. This will remove unincorporated liposomes.
#Suspend beads in 200-400ml desired buffer or in ddH2O. 

===== Laxex Bead Preparation: =====

#Measure out 10ml latex beads of the desired diameter.
#Wash 2X in 50mM carbonate-bicarbonate buffer, pH 9.6. Pellet cells with a max-speed spin in minifuge, 10min.
#Incubate 2hrs at 37oC, rotating constantly, with lipisome fractions prepared in “lipid preparation”.
#Wash 2X with carbonate-bicarbonate buffer
#Incubate with 5% BSA in PBS for 2+ hours to block beads. 

= Recipes =

===== Carbonate-bicarbonate buffer: =====

*1.15g Anhydrous sodium carbonate
*4.13g Sodium bicarbonate
*100ml ddH2O Dissolve in water, pH to 9.6.

Lipid Coated Bead Preparation - Revision history

Admin: Created page with "= Protocol = ===== Lipid Preparation: ===== #In a glass container, and using lipids dissolved in chloroform, mix desired lipids to a final quantity of ~4mmol total lipid. #..."