A procedure for the simultaneous determination of lipid and protein in biomembranes and other biological samples.
Very small sample sizes frequently become the limiting factor in biochemical and biomembrane studies in which routine quantification of protein and bulk lipids are required. The procedure described here allows the simultaneous determination of protein and lipid without initial, multiple aliquots. The method is based on the quantitative precipitation of proteins from a defined hexane/isopropanol mixture. The liquid phase resulting after decanting and concentrating to dryness can then be used to assay the lipid content directly. Quantitative assay of protein can be achieved after resuspension of the pelleted material by addition of sodium dodecyl sulfate (0.1%) and deoxycholate (1%). The method is also applicable to other types of lipid- and protein-containing samples with a broad range of protein/lipid ratios and lipid compositions, as they occur, for example, in serum lipoproteins.
Rodríguez-Vico
F
F
Department of Biochemistry and Molecular Biology, University of Granada, Spain.
Martínez-Cayuela
M
M
Zafra
M F
MF
García-Peregrin
E
E
Ramírez
H
H
D000433:1-Propanol; D000818:Animals; D002462:Cell Membrane / Q000737:chemistry*; D011232:Chemical Precipitation; D002645:Chickens; D002784:Cholesterol / Q000032:analysis; D006586:Hexanes; D008055:Lipids / Q000032:analysis*; D008074:Lipoproteins / Q000097:blood; D008297:Male; D008563:Membrane Lipids / Q000032:analysis; D008565:Membrane Proteins / Q000032:analysis; D008862:Microsomes, Liver / Q000737:chemistry; D010743:Phospholipids / Q000032:analysis; D011506:Proteins / Q000032:analysis*
D006586:Hexanes; D008055:Lipids; D008074:Lipoproteins; D008563:Membrane Lipids; D008565:Membrane Proteins; D010743:Phospholipids; D011506:Proteins; D000433:1-Propanol; D002784:Cholesterol