separated by funnel after adding ethyl ether (20 mL; Sigma-Aldrich, Saint Louis, MO, USA) was evaporated.
The collected infranatant
fraction is then brought back to its original volume, and the cholesterol content is then measured by the Abell-Kendall method.
At the end of the incubation period, an aliquot of the infranatant
was removed for enzymatic determination of glycerol released into the incubation medium (KATAL, Belo Horizonte, MG, Brazil).
fraction can now be reconstituted to the original volume with 0.
Therefore, the analysis of the clear infranatant
solution after ultracentrifugation will not give accurate triglyceride results.
We calculated the LDL-C concentration as the difference in the infranatant
between TC and HDL-C (see online Supplemental Data).
containing immature adipocytes was discarded and then the remained mature adipocytes were collected.
006 kg/L and removal of the supernatant containing chylomicrons and VLDL, apoB (designated LDL apoB) was measured in the infranatant
using 3 commercially available reagents each with antibodies to different apoB epitopes on automated chemistry analyzers.
To do this, we used sequential flotation ultracentrifugation for stepwise isolation of highly purified lipoprotein fractions (Athens Research and Technology), as described previously (6), with the exception that we also prepared IDL and lipoprotein particle-free infranatant
006 kg/L by ultracentrifugation (18 h; 105 000g; 4[degrees]C), and HDL-C was measured in the infranatant
by the direct method (instead of precipitation).
006 kg/L by ultracentrifugation (at 105 OOOg for 18 h at 4 [degrees]C) and measuring HDLc after precipitation in the infranatant
with phosphotungstate/ Mg[Cl.
LDLC was calculated as the difference between cholesterol in the infranatant
before and after precipitation, measured as described above.