BODIPY-Cholesterol
(Synonyms: BCh2) 目录号 : GC42964
A fluorescently tagged cholesterol
Cas No.:878557-19-8
Sample solution is provided at 25 µL, 10mM.
Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
| Cell experiment [1]: | |
|
Cell lines |
Human primary fibroblasts |
|
Preparation Method |
NPC1‐deficient GM3123 fibroblasts were incubated with Rhodamine‐dextran overnight to label terminal endocytic compartments, followed by incubation with 5 μg/mL (8.7 μm) BODIPY-Cholesterol in normal growth medium containing 5% fetal calf serum for 4 h, or 0.5 μm BODIPY-Cholesterol in medium containing 5% lipoprotein‐starved serum for 23 h. |
|
Reaction Conditions |
5 μg/mL, 4h |
|
Applications |
Time‐dependent partitioning of BODIPY-Cholesterol in the lysosomes of human primary fibroblasts. |
| Cell experiment [2]: | |
|
Cell lines |
NIH 3T3 |
|
Preparation Method |
NIH 3T3 were cultured in 24-well plates, incubated for 2 h with 2.5 µM BODIPY-cholesterol, and rinsed. Physiological buffer with or without 30 nM ShhN was added to the wells, and the BODIPY fluorescence intensity of the supernatants was measured after 1 h. |
|
Reaction Conditions |
2.5 µM; 2 h |
|
Applications |
The BODIPY fluorescence measured in the supernatants after 1 h was significantly lower in wells where ShhN protein was added compared to wells that did not contain ShhN. |
|
References: [1]. Hölttä-Vuori M, et al. Use of BODIPY-Cholesterol (TF-Chol) for Visualizing Lysosomal Cholesterol Accumulation. Traffic. 2016 Sep;17(9):1054-7. [2]. Bidet M, et al. The hedgehog receptor patched is involved in cholesterol transport. PLoS One. 2011;6(9):e23834. |
|
BODIPY-Cholesterol is cholesterol tagged with a boron dipyrromethene difluoride (BODIPY) fluorophore used for monitoring sterol uptake and inter-organelle sterol flux in cells with excitation of 480 nm and emission of 508 nm.[1] The excretion of BODIPY cholesterol from late endoplasmic organelles depends on acidic lipase and Niemann pickc1 protein.[6]
In vitro, treatment with Bodipy-cholesterol in cells, prominent PM labeling is observed at 2–5 min; however, upon ≥30 min incubations, it is also observed the fluorescence labeling of intracellular structures.[2] In vitro efficacy test it suggested that accumulation of BODIPY-cholesterol in the media gave more reproducible values than assaying for the loss of the compound from the cells.[3] With 1 μg BODIPY-cholesterol analogs show a similar cellular localization in HeLa cells and exhibit similar cholesterol efflux properties from THP1 cells to HDL acceptors.[4] BODIPY-cholesterol efflux clearly increased when treatment of fibroblasts with the Hh pathway agonist SAG, which enhances Ptc protein expression, or over-expression of human Ptc in yeast.[5] Treatment with miR-758 inhibitor obviously increased ABCA1-dependent cholesterol efflux by BODIPY-Cholesterol efflux assay.[7] Micrographs of EPCs incubated with HDL labeld bodipy-cholesterol (50 μg/ml) 30 min, small cytoplasmic vesicles as well as large positive MVBs containing intraluminal microvesicles, tightly-packed with reaction products could be displayed. The internalized-HDL-derived bodipy-cholesterol was also spread within many of the stacked Golgi cisterns and the TGN.[8]
References:
[1].Wüstner D, et al. Potential of BODIPY-cholesterol for analysis of cholesterol transport and diffusion in living cells. Chem Phys Lipids. 2016 Jan;194:12-28.
[2].H?ltt?-Vuori M, et al. BODIPY-cholesterol: a new tool to visualize sterol trafficking in living cells and organisms. Traffic. 2008 Nov;9(11):1839-49.
[3].Sankaranarayanan S, et al. A sensitive assay for ABCA1-mediated cholesterol efflux using BODIPY-cholesterol. J Lipid Res. 2011 Dec;52(12):2332-2340.
[4].Liu Z, et al. Synthesis of cholesterol analogues bearing BODIPY fluorophores by Suzuki or Liebeskind-Srogl cross-coupling and evaluation of their potential for visualization of cholesterol pools. Chembiochem. 2014 Sep 22;15(14):2087-96.
[5].Bidet M, et al. The hedgehog receptor patched is involved in cholesterol transport. PLoS One. 2011;6(9):e23834.
[6].Kanerva K, et al. LDL cholesterol recycles to the plasma membrane via a Rab8a-Myosin5b-actin-dependent membrane transport route. Dev Cell. 2013 Nov 11;27(3):249-62.
[7].Yao Y, et al. Glucagon-like peptide-1 contributes to increases ABCA1 expression by downregulating miR-758 to regulate cholesterol homeostasis. Biochem Biophys Res Commun. 2018 Mar 4;497(2):652-658.
[8].Srisen K, et al. Human endothelial progenitor cells internalize high-density lipoprotein. PLoS One. 2013 Dec 30;8(12):e83189.
BODIPY-Cholesterol 是用硼二氟甲烷 (BODIPY) 荧光团标记的胆固醇，用于监测细胞中甾醇摄取和细胞器间甾醇通量，激发波长为 480 nm，发射波长为 508 nm。[1] 晚期内质细胞器排泄 BODIPY 胆固醇依赖于酸性脂肪酶和 Niemann pickc1 蛋白。[6]
在体外，在细胞中用 Bodipy-cholesterol 处理，在 2-5 分钟时观察到显着的 PM 标记；然而，在≥30分钟的孵育后，也观察到细胞内结构的荧光标记。[2]体外功效测试表明，BODIPY-胆固醇在培养基中的积累比测定具有更高的可重复性值[3]1 μg BODIPY-胆固醇类似物在 HeLa 细胞中表现出相似的细胞定位，并表现出相似的胆固醇从 THP1 细胞流出到 HDL 受体的特性。[4]当用 Hh 通路激动剂 SAG 处理成纤维细胞时，BODIPY-胆固醇流出明显增加，这会增强 Ptc 蛋白表达，或人 Ptc 在酵母中的过度表达。[5] 通过 BODIPY-Cholesterol 流出测定，用 miR-758 抑制剂处理明显增加了 ABCA1 依赖性胆固醇流出。[7] EPCs 显微照片与 HDL 标记的 bodipy-cholesterol (50 μg/ml) 孵育 30 分钟，小细胞质囊泡以及大的阳性 MVBs 含有内可以显示与反应产物紧密堆积的管腔微泡。内化 HDL 衍生的身体胆固醇也分布在许多堆叠的高尔基池和 TGN 中。[8]
| Cas No. | 878557-19-8 | SDF | |
| 别名 | BCh2 | ||
| 化学名 | (T-4)-[(3β)-24-(3,5-dimethyl-1H-pyrrol-2-yl-κN)-24-(3,5-dimethyl-2H-pyrrol-2-ylidene-κN)chol-5-en-3-olato]difluoro-boron | ||
| Canonical SMILES | O[C@H](C1)CC[C@@]2(C)C1=CC[C@]3([H])[C@]2([H])CC[C@@]4(C)[C@@]3([H])CC[C@]4([H])[C@H](C)CCC(C5=C(C)C=C(C)N5[B-]6(F)F)=C7[N+]6=C(C)C=C7C | ||
| 分子式 | C36H51BF2N2O | 分子量 | 576.6 |
| 溶解度 | 0.5mg/ml in ethanol; 1mg/ml in DMSO, or in DMF | 储存条件 | Store at -20°C |
| General tips | 请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
||
| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
 |
1 mg | 5 mg | 10 mg |
| 1 mM | 1.7343 mL | 8.6715 mL | 17.343 mL |
| 5 mM | 0.3469 mL | 1.7343 mL | 3.4686 mL |
| 10 mM | 0.1734 mL | 0.8672 mL | 1.7343 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
Potential of BODIPY-Cholesterol for analysis of cholesterol transport and diffusion in living cells
Chem Phys Lipids 2016 Jan;194:12-28.PMID:26291493DOI:10.1016/j.chemphyslip.2015.08.007.
Cholesterol is an abundant and important lipid component of cellular membranes. Analysis of cholesterol transport and diffusion in living cells is hampered by the technical challenge of designing suitable cholesterol probes which can be detected for example by optical microscopy. One strategy is to use intrinsically fluorescent sterols, as dehydroergosterol (DHE), having minimal chemical alteration compared to cholesterol but giving low fluorescence signals in the UV region of the spectrum. Alternatively, one can use dye-tagged cholesterol analogs and in particular BODIPY-Cholesterol (BChol), whose synthesis and initial characterization was pioneered by Robert Bittman. Here, we give a general overview of the properties and applications but also limitations of BODIPY-tagged cholesterol probes for analyzing intracellular cholesterol trafficking. We describe our own experiences and collaborative efforts with Bob Bittman for studying diffusion in the plasma membrane (PM) and uptake of BChol in a quantitative manner. For that purpose, we used a variety of fluorescence approaches including fluorescence correlation spectroscopy and its imaging variants, fluorescence recovery after photobleaching (FRAP) and fluorescence loss in photobleaching (FLIP). We also describe pulse-chase studies from the PM using BChol in direct comparison to DHE. Based on the gathered imaging data, we present a two-step kinetic model for sterol transport between PM and recycling endosomes. In addition, we highlight the suitability of BChol for determining transport of lipoprotein-derived sterol using electron microscopy (EM) and show that this approach ideally complements fluorescence studies.
Use of Bodipy-labeled sphingolipid and cholesterol analogs to examine membrane microdomains in cells
Histochem Cell Biol 2008 Nov;130(5):819-32.PMID:18820942DOI:10.1007/s00418-008-0509-5.
Much evidence has accumulated to show that cellular membranes such as the plasma membrane, contain multiple "microdomains" of differing lipid and protein composition and function. These domains are sometimes enriched in cholesterol and sphingolipids and are believed to be important structures for the regulation of many biological and pathological processes. This review focuses on the use of fluorescent (Bodipy) labeled analogs of sphingolipids and cholesterol to study such domains. We discuss the similarities between the behavior of BODIPY-Cholesterol and natural cholesterol in artificial bilayers and in cultured cells, and the use of Bodipy-sphingolipid analogs to visualize membrane domains in living cells based on the concentration-dependent monomer-excimer fluorescence properties of the Bodipy-fluorophore. The use of Bodipy-D-erythro-lactosylceramide is highlighted for detection of domains on the plasma membrane and endosome membranes, and the importance of the sphingolipid stereochemistry in modulating domain formation is discussed. Finally, we suggest that Bodipy-sphingolipids may be useful in future studies to examine the relationship between membrane domains at the cell surface and domains enriched in other lipids and proteins on the inner leaflet of the plasma membrane.
BODIPY-Cholesterol can be reliably used to monitor cholesterol efflux from capacitating mammalian spermatozoa
Sci Rep 2019 Jul 8;9(1):9804.PMID:31285440DOI:10.1038/s41598-019-45831-7.
Capacitation is the final maturation step spermatozoa undergo prior to fertilisation. The efflux of cholesterol from the sperm membrane to the extracellular environment is a crucial step during capacitation but current methods to quantify this process are suboptimal. In this study, we validate the use of a BODIPY-Cholesterol assay to quantify cholesterol efflux from spermatozoa during in vitro capacitation, using the boar as a model species. The novel flow cytometric BODIPY-Cholesterol assay was validated with endogenous cholesterol loss as measured by mass spectrometry and compared to filipin labelling. Following exposure to a range of conditions, the BODIPY-Cholesterol assay was able to detect and quantify cholesterol efflux akin to that measured with mass spectrometry. The ability to counterstain for viability is a unique feature of this assay that allowed us to highlight the importance of isolating viable cells only for a reliable measure of cholesterol efflux. Finally, the BODIPY-Cholesterol assay proved to be the superior method to quantify cholesterol efflux relative to filipin labelling, though filipin remains useful for assessing cholesterol redistribution. Taken together, the BODIPY-Cholesterol assay is a simple, inexpensive and reliable flow cytometric method for the measurement of cholesterol efflux from spermatozoa during in vitro capacitation.
Liraglutide improves lipid metabolism by enhancing cholesterol efflux associated with ABCA1 and ERK1/2 pathway
Cardiovasc Diabetol 2019 Nov 9;18(1):146.PMID:31706303DOI:10.1186/s12933-019-0954-6.
Background: Reverse cholesterol transport (RCT) is an important cardioprotective mechanism and the decrease in cholesterol efflux can result in the dyslipidemia. Although liraglutide, a glucagon like peptide-1 analogue, has mainly impacted blood glucose, recent data has also suggested a beneficial effect on blood lipid. However, the exact mechanism by which liraglutide modulates lipid metabolism, especially its effect on RCT, remain undetermined. Hence, the aim of the present study was to investigate the potential impacts and potential underlying mechanisms of liraglutide on the cholesterol efflux in both db/db mice and HepG2 cells. Methods: Six-week old db/db mice with high fat diet (HFD) and wild type mice were administered either liraglutide (200 μg/kg) or equivoluminal saline subcutaneously, twice daily for 8 weeks and body weight was measured every week. After the 8-week treatment, the blood was collected for lipid evaluation and liver was obtained from the mice for hematoxylin-eosin (HE) staining, red O staining and Western blotting. Cholesterol efflux was assessed by measuring the radioactivity in the plasma and feces after intraperitoneal injection of 3H-labeled cholesterol. HepG2 Cells were treated with different concentrations of glucose (0, 5, 25, and 50 mmol/L) with or without liraglutide (1000 nmol/L) for 24 h. The intracellular cholesterol efflux was detected by BODIPY-Cholesterol fluorescence labeling. Real-time PCR or Western blotting was used to examine the expression levels of ABCA1, ABCG1 and SR-B1. Results: Liraglutide significantly decreased blood glucose, serum total cholesterol (TC), triglyceride (TG) and low-density lipoprotein cholesterol (LDL-C). It also reduced liver lipid deposition in db/db mice fed with HFD. Moreover, the movement of 3H-cholesterol from macrophages to plasma and feces was significantly enhanced in db/db mice fed with HFD after liraglutide adminstration. In vitro study, liraglutide could promote the cholesterol efflux of HepG2 cells under high glucose, and also increase the expression of ABCA1 by activating the ERK1/2 pathway. Conclusions: Liraglutide could improve lipid metabolism and hepatic lipid accumulation in db/db mice fed with HFD by promoting reversal of cholesterol transport, which was associated with the up-regulation of ABCA1 mediated by the ERK1/2 phosphorylation.
An assay to evaluate the capacity of cholesterol acceptors using BODIPY-Cholesterol in cells
STAR Protoc 2023 Jan 3;4(1):101976.PMID:36598853DOI:10.1016/j.xpro.2022.101976.
Cholesterol is a structural component of cell membranes. Most cells are incapable of its catabolism, and intracellular cholesterol accumulation is linked to several disorders including cardiovascular and neurodegenerative diseases. Cholesterol efflux, essential to its metabolism, is dependent on acceptors such as apolipoproteins. Here, we describe an assay to evaluate the capacity of cholesterol acceptors. Cells are treated with an analog of cholesterol tagged with fluorescent BODIPY. Addition of an acceptor leads to BODIPY-Cholesterol efflux, measured using a plate reader. For complete details on the use and execution of this protocol, please refer to Liu et al. (2021).1.