Separation of serum lipoproteins of Japanese quail by disc polyacrylamide gel electrophoresis and single discontinuous density gradient ultracentrifugation.
A method based on disc polyacrylamide gel electrophoresis (disc PAGE) for the separation of serum lipoproteins of Japanese quail (Coturnix coturnix japonica) prestained with Sudan Black B is described and evaluated. Good separation was obtained by using 3% separating gel and decreasing the concentration of buffer solution A used for preparation of separating gel solution to one-half that recommended by Narayan (1975). Best resolution of the high density lipoproteins was achieved by using 7 to 10% of separating gel, which separated the high density lipoproteins into three bands. Clear lipoprotein bands were obtained after ultracentrifugation of prestained serum. For corresponding positions, the bands of unstained serum after staining had the same mobility in the disc (PAGE) as prestained lipoprotein bands. In quail, density smaller than 1.006 g/ml contains chylomicrons and very low density lipoproteins (VLDL); density from 1.018 to 1.05 g/ml contains the low density lipoprotein (LDL); and density more than 1.05 g/ml and less than 1.16 g/ml contains the high density lipoproteins (HDL). In the profile of serum lipoproteins after ultracentrifugation, two peaks were observed in the density range of HDL. There was one small peak with density 1.05 to 1.09 g/ml and one large peak with density 1.09 to 1.16 g/ml. Based on the lipoprotein profiles from both disc PAGE and ultracentrifugation, HDL is the predominant form, LDL is intermediate, and VLDL and chylomicrons are smallest in amount.
D000818:Animals; D002499:Centrifugation, Density Gradient; D002914:Chylomicrons / Q000097:blood; D003370:Coturnix / Q000097:blood*; D004591:Electrophoresis, Polyacrylamide Gel; D008074:Lipoproteins / Q000097:blood*; D008075:Lipoproteins, HDL / Q000097:blood; D008077:Lipoproteins, LDL / Q000097:blood; D008079:Lipoproteins, VLDL / Q000097:blood; D008297:Male; D011784:Quail / Q000097:blood*
D002914:Chylomicrons; D008074:Lipoproteins; D008075:Lipoproteins, HDL; D008077:Lipoproteins, LDL; D008079:Lipoproteins, VLDL