https://wiki.phagocytes.ca/index.php?action=history&feed=atom&title=Lipid_Extraction_from_Cells 2026-04-28T14:51:19Z Revision history for this page on the wiki MediaWiki 1.40.1 https://wiki.phagocytes.ca/index.php?title=Lipid_Extraction_from_Cells&diff=72&oldid=prev 2021-02-01T19:58:36Z

<p>Created page with &quot;= Protocol = ===== Initial extraction: ===== #Wash cells 3X with PBS to remove serum. #Remove cells by scraping cells into PBS. #Pellet cells in ultra centrifuge; max spee...&quot;</p> <p><b>New page</b></p><div>= Protocol =<br /> <br /> ===== Initial extraction: =====<br /> <br /> #Wash cells 3X with PBS to remove serum. <br /> #Remove cells by scraping cells into PBS. <br /> #Pellet cells in ultra centrifuge; max speed for 5min. <br /> #Add ~20X volume of 1:1 chloroform:methanol, vortex to suspend and rotate 20min at RT. <br /> #Spin pellet max speed for 5min, recover fluid phase.&lt;br&gt; <br /> <br /> ===== Option 1: Crude extract alone =====<br /> <br /> #Dry chloroform solution from step 5 above, using inert gas (N2, CO2, argon, etc). <br /> #Re-suspend lipids in desired media or solvent. <br /> <br /> *Note: this solution will contain gangliosides, small polar molecules, etc.&lt;br&gt; <br /> <br /> ===== Option 2: Lipid Purification =====<br /> <br /> #Mix a 0.2 volume of PBS into the mixture produced by step 5 of “Initial extraction”, vortex several seconds to mix. <br /> #Centrifuge 2000rpm in minifuge to separate phases. Upper (water) phase contains gangliosides, lower (chloroform) layer contains purified lipids. <br /> #Dry the lower layer under inert gas <br /> #Suspend as needed</div>

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