반응 #669405

반응 방정식

SDS

glucose

cholesterol

25-hydroxycholesterol

→

cholesteryl oleate

반응 조건

후처리

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

실험 절차

More specifically, cultured J774 cells were plated on a 24 F culture plate containing a serum-free RPMI 1640 medium so as to account for 1×10-6 cells/well. Added to the cells were [14C]oleic acid, delipidized BAS (bovine serum albumin), reconstituted ribosome (0.3 M 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 dodecyl 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.

출처