BODIPY-cholesterol

Laboratory Procedures

BODIPY-cholesterol is supplied as a crystalline solid.
BODIPY-cholesterol作为晶体固体提供。
A stock solution may be made by dissolving the BODIPY-cholesterol in the solvent of choice.
储备溶液可以通过在所选择的溶剂中溶解身体的胆固醇来制备。
BODIPY-cholesterol is soluble in organic solvents such as ethanol, DMSO, and dimethyl formamide(DMF), which should be purged with an inert gas.
BODIPY-cholesterol溶于有机溶剂，如乙醇、DMSO和二甲基甲酰胺（DMF），这些溶剂应该用惰性气体净化。
The solubility of BODIPY-cholesterol in ethanol is approximately 0.5 mg/ml and approximately 1 mg/ml in DMSO and DMF.
BODIPY-cholesterol在乙醇中的溶解度约为0.5 mg/ml，在DMSO和DMF中的溶解度约为1 mg/ml。
BODIPY-cholesterol is sparingly soluble in aqueous buffers.
BODIPY-cholesterol在水缓冲液中可溶。
For maximum solubility in aqueous buffers, BODIPY-cholesterol should first be dissolved in DMSO and then diluted with the aqueous buffer of choice BODIPY-cholesterol has a solubility of approximately 0.5 mg/ml in a 1: 1 solution of DMSO: PBS(pH 7. 2) using this method.
为了在水缓冲液中获得最大溶解度，BODIPY-cholesterol应首先溶解在DMSO中，然后用所选的水缓冲液稀释，BODIPY-cholesterol在DMSO:PBS（pH 7.2）为1:1溶液中的溶解度约为0.5 mg/ml。
We do not recommend storing the aqueous solution for more than one day.
用这种方法。我们不建议将水溶液保存超过一天。

Description

BODIPY-cholesterol is a biologically active and cell-permeable analog of cholesterol that is tagged with a fluorescent BODIPY group at carbon 24.1.2
BODIPY-cholesterol是一种具有生物活性和细胞通透性的胆固醇类似物，在碳24处标记有荧光BODIPY基团。
It co-localizes with dehydroergosterol, a marker of cholesterol, in HeLa cells and is trafficked from the plasma membrane to the endocytic recycling compartment in BHK cells. 2
它与胆固醇标记物脱氢麦角甾醇在HeLa细胞中共定位，并从BHK细胞质膜运输到的内吞循环室。2
BODIPY-cholesterol displays excitation/emission maxima of 480/508 nm, respectively, and has been used to monitor sterol uptake and inter-organelle sterol flux in cells
BODIPY-cholesterol显示最大激发/发射波长分别为480/508nm，并被用于监测细胞内固醇摄取和细胞器间固醇通量。

BODIPY-cholesterol efflux assay

BODIPY-cholesterol流出试验

BODIPY-cholesterol (Glpbio, America) solutions were prepared in DMSO at a concentration of 5 mM.
用DMSO制备了BODIPY-cholesterol（Glpbio，美国）为5 mM储存液。
VSMCs were cultured in 6-well plates and labeled with 2.5 mM of BODIPY-cholesterol in culture medium for 2 h at 37 °C followed by washing with MEM-HEPES(Gibco, America).
在6孔培养板中培养VSMCs 细胞，在37℃，用2.5 mM的BODIPY-cholesterol 标记2 h，然后用MEM-HEPES（Gibco，美国）洗涤。
After that, the efflux medium was removed and filtered through a 0.45 μm filter. The BODIPY fluorescence intensity was examined by using a fluorescence photometer (excitation 490 ± 10 nm; emission 520 ± 20 nm).
之后，除去流出培养基，并通过0.45μm过滤器进行过滤。用荧光光度计（激发490    10 nm；发射520   20 nm）检测BODIPY荧光强度。
For fluorescence microscopy observations, VSMCs were cultured on 12-well plate containing 2.5 mM BODIPY-cholesterol for 2 h at 37 °C, and then rinsed with MEM-HEPES.
荧光显微镜观察：VSMCs在含有2.5 mM BODIPY-cholesterol 的12孔板上37°C培养2 h，然后用MEM-HEPES冲洗。
Cells were fixed with 4% paraformaldehyde for 10 min after rinsed with PBS, and observed using the fluorescence microscope. BODIPY fluorescence was analyzed by using Image J software.
用PBS冲洗细胞后，用4%多聚甲醛固定10 min，用荧光显微镜观察。用Image J软件对BODIPY荧光进行了分析。