Wako l type ldl cholesterol

After fasting overnight, mice were anesthetized briefly with 5% isoflurane (Abbott Animal Health) inhalation and 200 μl of blood were collected by retro-orbital plexus using a noncoated hematocrit glass capillary (Chase Scientific Glass). Blood was allowed to clot at 4°C overnight and then centrifuged at 5,000 g for 15 min (4°C). The obtained supernatant was centrifuged again at 11,000 g for 2 min (4°C). Serum total cholesterol, LDL-C, and HDL-C were measured using commercial colorimetric assays (Wako Cholesterol E, Wako L-type LDL-Cholesterol, Wako HDL-Cholesterol E; Wako Diagnostics) according to the manufacturer’s instructions.

After fasting overnight, mice were anesthetized briefly with 5% isoflurane (Abbott Animal Health, Abbott Park, IL) inhalation and 200 μl of blood were collected by retro-orbital plexus using a noncoated hematocrit glass capillary (Chase Scientific Glass, Rockwood, TN). Blood was allowed to clot at 4°C overnight and then centrifuged at 5,000 g for 15 min (4°C). The obtained supernatant was centrifuged again at 11,000 g for 2 min (4°C). Serum total cholesterol, LDL-cholesterol (LDL-C), and HDL-C were measured using commercial colorimetric assays (Wako Cholesterol E, Wako L-type LDL-Cholesterol, Wako HDL-Cholesterol E; Wako Diagnostics, Richmond, VA) according to the manufacturer’s instructions. HDL-C levels were measured in plasma from bone marrow (BM)-transplanted mice by using HDL and LDL/VLDL quantification kit (Sigma-Aldrich) following the manufacturer’s instructions. Four volumes of blood were collected from the inferior vena cava in a tube containing 1 vol of 3.2% sodium citrate. Blood was centrifuged at 5,000 g for 15 min at room temperature and then the supernatant was centrifuged again at 11,000 g for 3 min at room temperature. Plasma was stored at −80°C until use.