An acetylation method for the quantification of membrane lipids, including phospholipids, polyphosphoinositides and cholesterol.
A method for the quantification of membrane lipids by acetylation with [3H]acetic anhydride is described. A standard lipid, labelled with 14C or 32P, is added to the sample and is simultaneously acetylated. The 3H/14C or 3H/32P ratios obtained in the acetylated lipids are proportional to the initial lipid concentration. The method has been used to quantify cholesterol, phosphatidylethanolamine, phosphatidylinositol, sphingomyelin, phosphatidylinositol 4-phosphate and phosphatidylinositol 4,5-bisphosphate in amounts between 2 and 10 nmol.
Stein
J M
JM
Department of Clinical Biochemistry, University of Cambridge, Addenbrooke's Hospital, U.K.
Smith
G A
GA
Luzio
J P
JP
D000107:Acetylation; D002250:Carbon Radioisotopes; D002458:Cell Fractionation; D002462:Cell Membrane / Q000737:chemistry* / Q000648:ultrastructure; D002499:Centrifugation, Density Gradient; D002784:Cholesterol / Q000032:analysis* / Q000097:blood; D002855:Chromatography, Thin Layer / Q000379:methods; D003124:Colorimetry / Q000379:methods; D006801:Humans; D007202:Indicators and Reagents; D008563:Membrane Lipids / Q000032:analysis* / Q000097:blood; D009504:Neutrophils / Q000737:chemistry*; D018129:Phosphatidylinositol Phosphates; D010716:Phosphatidylinositols / Q000032:analysis* / Q000097:blood; D010743:Phospholipids / Q000032:analysis* / Q000097:blood; D010761:Phosphorus Radioisotopes; D011865:Radioisotope Dilution Technique; D014316:Tritium
D002250:Carbon Radioisotopes; D007202:Indicators and Reagents; D008563:Membrane Lipids; D018129:Phosphatidylinositol Phosphates; D010716:Phosphatidylinositols; D010743:Phospholipids; D010761:Phosphorus Radioisotopes; D014316:Tritium; D002784:Cholesterol