pH gradient electrophoresis and isoelectric focusing of lipoproteins on agarose bead thin layers.
A new method of isoelectric focusing (IEF) and pH gradient electrophoresis, using thin layers of agarose gel beads, was devised to investigate chylomicrons and very low density lipoproteins (VLDL). pH gradient stability and cathodal gradient drift were similar to those of polyacrylamide gel IEF, and linearity of gradients was maintained for 23 hr. Chylomicrons and VLDL were detectable without staining. Chylomicrons from human serum and from rat lymph migrated in this system. Rat lymph chylomicrons, obtained by ultracentrifugation, migrated in several discrete bands, and this heterogeneity or rat chylomicrons was confirmed by electron microscopic demonstration of chylomicrons in each band. This new technique has permitted the first measurement of isoelectric points of some lipoproteins in the ultracentrifuged fraction of human serum chylomicrons and the first separation of multiple discrete fractions of ultracentrifuged lymph chylomicrons.
D000818:Animals; D002855:Chromatography, Thin Layer / Q000379:methods; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing / Q000379:methods; D008074:Lipoproteins / Q000097:blood / Q000302:isolation & purification*; D008196:Lymph / Q000032:analysis; D008297:Male; D051381:Rats; D012685:Sepharose
D008074:Lipoproteins; D012685:Sepharose