The blood samples were collected into serum-separating tubes. After a brief centrifugation (2000× g, 30 min) to remove the residual debris of blood cells, the supernatant was prepared for LPEP by isoelectric focusing methods using a commercial kit (Quick gel, Helena Laboratories, Saitama, Japan). Briefly, 30 μL of each blood sample was loaded onto the agarose gel for migration at 250V and 20 °C. Lipoproteins in the blood samples were separated into four major bands on agarose gels based on their electric charges: chylomicron, VLDL (pre-β lipoprotein), LDL (β-lipoprotein), and HDL (α-lipoprotein) [14 (link)]. Each band was assessed and analyzed by densitometric scanning by using Edbank III analysis software (Helena Laboratories, Saitama, Japan) to determine the proportion of each lipoprotein fraction. The presence of chylomicrons in the fasting blood determined by LPEP was an important characteristic feature of HLP5, whereas chylomicrons in the fasting blood was absent in HLP4 [1 (link),5 (link),6 (link)].
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