Whole lipoproteins from serum of a healthy person were analyzed using PROTEIN KW-804, an aqueous SEC (GFC) column. The result shows a good separation of the three kinds of lipoproteins (VLDL, LDL and HDL) in serum.
(Preparation of sample) - Adjust specific gravity of serum of a healthy person to 1.210g/mL added potassium bromide followed by ultracentrifugation for 24 hours. - After dialyzing the supernatant, solvent substitute by PBS*. - Measure protein concentration by Lowry method and dilute with PBS* to 1.0mg/mL.
Sample : 40µL Whole lipoproteins from serum of a healthy person 1.0mg/mL 1. VLDL 2. LDL 3. HDL
Columns : Shodex PROTEIN KW-G 6B (6.0mmI.D. x 50mm) + KW-804 (8.0mmI.D. x 300mm)
Eluent : 10-fold diluted x10 PBS* with H2O
Flow rate : 1.0mL/min
Detector : UV(280nm)
Column temp. : 30°C
x10 PBS* : 80g NaCl + 29g Na2HPO4・12H2O + 2g KCl + 2g KH2PO4 in 1000mL of H2O
Data courtesy of Ohkawa Ryunosuke, Graduate School of Health Care Sciences, Analytical Laboratory Chemistry, Tokyo Medical and Dental University