Inositol Labeling and Lipid Extraction: Difference between revisions

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 ==Materials==
-�*Cells in 100 mm culture dishes
+*Cells in 100 mm culture dishes
-�*Labelling Media (Inositol Free DMEM with PSG, 10% dialysed serum and 100 uCi inositol per plate
+*Labelling Media (Inositol Free DMEM with PSG, 10% dialysed serum and 100 uCi inositol per plate
 of cells)
-�*Perchloric acid (640 uL into 10 mL water) keep cold
+*Perchloric acid (640 uL into 10 mL water) keep cold
-�*100 mM EDTA (2 mL of 0.5M stock in 10 mL) keep cold
+*100 mM EDTA (2 mL of 0.5M stock in 10 mL) keep cold
-�*Water
+*Water
-�*Deacylation Mix (3.5 mL Methylamine, 6 mL Methanol, 1.5 mL Butanol, 2.1 mL Water)
+*Deacylation Mix (3.5 mL Methylamine, 6 mL Methanol, 1.5 mL Butanol, 2.1 mL Water)
-�*Lipid Extraction Mix (10 mL Butanol, 2 mL Ethyl Ether and 0.5 mL Ethyl Formate)
+*Lipid Extraction Mix (10 mL Butanol, 2 mL Ethyl Ether and 0.5 mL Ethyl Formate)
 ==Protocol==