反应 #448877

反应方程式

cholesterol

25-hydroxycholesterol

SDS

glucose

→

cholesteryl oleate

反应条件

后处理

1. 1
   workup.ADDITIONcontaining a serum-free RPMI 1640 medium so as
2. 2
   workup.ADDITIONAdded to the cells
3. 3
   萃取the lipid in the disrupted solution was extracted with hexane
4. 4
   其他The extract was dried to solid under reduced pressure

实验过程

More specifically, cultured J774 cells were plated on a 24F culture plate containing a serum-free RPMI 1640 medium so as to account for 1×106 cells/well. Added to the cells were [14C]oleic acid, delipidized BAS (bovine serum albumin), reconstituted ribosome (0.3M glucose solution containing cholesterol and phosphatidylcholine at a weight ratio of 2:1) and 25-hydroxycholesterol to culture the cells for 4 hours at 37° C. under 5% CO2. After the culturing, the cultured cells were disrupted with a 1% SDS (sodium lauryl sulfate) solution, and the lipid in the disrupted solution was extracted with hexane. The extract was dried to solid under reduced pressure. After the resultant residue was then developed (developing solvent: diethyl ether/hexane/acetic acid=80/20/1) by TLC (thin-layer chromatography), the amount of cholesteryl oleate formed was determined by an imaging plate.

来源