Filipin III
(Synonyms: 菲律宾菌素 III) 目录号 : GC12048An isomeric polyene macrolide
Cas No.:480-49-9
Sample solution is provided at 25 µL, 10mM.
-
Related Biological Data
Tubeimosides inhibit NPC1. B SNB-19 cells were treated overnight with indicated compounds. After being fixed, cells were stained with filipin (50 µg/mL) and visualized by confocal microscopy (scale bar, 40 μm). Experiments were repeated 3 times independently, and representative results are shown.
After being fixed with 4% paraformaldehyde, cells were washed twice with PBS, and incubated with 50 µg/ml filipin III(GlpBio) in PBS for 1 h in dark at room temperature.
Nature Communications, 2024, 15(1): 162. PMID: 38167417 IF: 16.6009 -
Related Biological Data
24HC induces cholesterol accumulation in multivesicular bodies or late endosomes.D. N2a cells were treated with ETOH, 24HC (5 µM), or 25HC (5 µM) for 24 h and then fixed and stained with filipin (cyan) and Rab7 (red). Cells were analyzed for the co-localization between filipin and Rab7. White boxes indicate the magnified sections of the images.
For cholesterol staining, Filipin III (Glpbio) was used after fixing the cells.
Redox Biology (2023): 102769.. PMID: 37285742 IF: 10.7865 -
Related Biological Data
Studying the cell entry pathway of Antimir21-RNP-Apt. (a) The probability of observing a force–time signal for Antimir21-RNP-Apt entry into HEK-293 cells and MDA-MB-231 cells (before and after inhibition).
After adding the inhibitors namely EIPA (10 µM), nystatin (10 µg mL−1), filipin III (1 µg mL−1) (GlpBio), and CPZ (1 µg mL−1) and incubating for 30 min, the cells were washed with PBS and fresh medium was added to carry out the force tracing experiment.
Nanoscale 14.25 (2022): 8919-8928. PMID: 35699091 IF: 6.7
Quality Control & SDS
- View current batch:
- Purity: >90.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
本方案仅提供一个指导，请根据您的具体需要进行修改。
1.制备染色液
(1)配置DMSO储存液:使用DMSO或无水乙醇溶解Filipin III，配置浓度为1-5mM的储存液。
注意:未使用的储存液分装后在-20℃或-80°C避光保存，避免反复冻融。
(2)配置工作液:用合适的缓冲液(如:无血清培养基或PBS)稀释储存液，配制浓度为1-250μM的工作液。
注意:请根据实际情况调整工作液浓度，现用现配。
2.细胞悬浮染色
(1)悬浮细胞：经4°C、1000g离心3-5分钟，弃去上清液，用PBS清洗两次，每次5分钟。
(2)贴壁细胞：使用PBS清洗两次，加入胰酶消化细胞，消化完成后经1000g离心3-5min。
(3)加入Filipin III工作溶液重悬细胞，室温避光孵育15-30分钟。不同细胞最佳孵育时间不同，请根据具体实验需求自行摸索。
(4)孵育结束后，经1000g离心5分钟，去除上清液，加入PBS清洗2-3次，每次5分钟。
(5)用预温的无血清细胞培养基或PBS重悬细胞。通过荧光显微镜或流式细胞术观察。
3.细胞贴壁染色
(1)在无菌盖玻片上培养贴壁细胞。
(2)从培养基中移走盖玻片，吸出过量的培养基，将盖玻片放在潮湿的环境中。
(3)从盖玻片的一角加入100μL的染料工作液，轻轻晃动使染料均匀覆盖所有细胞。
(4)室温避光孵育15-30分钟。不同细胞最佳孵育时间不同，请根据具体实验需求自行摸索。
(5)孵育结束后吸弃染料工作液，使用预温的培养液清洗盖玻片2~3次。
4.显微镜检测：Filipin III的激发光范围是340-380nm，发射光范围是385-470 nm。
注意事项：
1）荧光染料均存在淬灭问题，请尽量注意避光，以减缓荧光淬灭。
2）为了您的安全和健康，请穿实验服并戴一次性手套操作。
Cholesterol is both an important structural component of cell membranes and an early intermediate in hormone and bile acid biosynthesis. The localization and measurement of cholesterol in cells is therefore of great interest. Filipin is the collective name given to four isomeric polyene macrolides isolated from cultures of S. filipinensis; Filipin III is the predominant isomer and the one used in most studies. Filipin binds to cholesterol in membranes, forming ultrastructural aggregates and complexes which can be visualized by freeze-fracture electron microscopy.[1],[2] The binding of cholesterol also decreases the intrinsic fluorescence of Filipin, and this property has also been used to detect cholesterol in membrane fractions.[3]
Reference:
[1]. Castanho, M.A., Coutinho, A., and Prieto, M.J. Absorption and fluorescence spectra of polyene antibiotics in the presence of cholesterol J. Biol. Chem. 267(1), 204-209 (1992).
[2]. Miller, R.G. Mini Review. The use and abuse of filipin to localize cholesterol in membranes Cell Biol. Int. Rep. 8(7), 519-535 (1984).
[3]. Severs, N.J., and Robenek, H. Detection of microdomains in biomembranes. An appraisal of recent developments in freeze-fracture cytochemistry Biochimica et Biophysica Acta 737, 373-408 (1983)
胆固醇既是细胞膜的重要结构成分，也是激素和胆汁酸生物合成的早期中间体。因此，在细胞中定位和测量胆固醇非常重要。菲洛平（Filipin）是从S. filipinensis培养物中分离出来的四种同分异构体多烯大环内酯的集合名称；菲洛平III是主要同分异构体，也是大多数研究所使用的一种。菲洛平与膜中的胆固醇结合，形成超微结构聚集体和复合物，可以通过冷冻切片电子显微镜观察到[1] [2]。 胆固醇的结合还会降低菲洛平本身荧光强度，并且这个特性也被用于检测薄膜部分中的胆固醇[3]。
| Cas No. | 480-49-9 | SDF | |
| 别名 | 菲律宾菌素 III | ||
| 化学名 | (3R,4S,6S,8S,10R,12R,14R,16S,17E,19E,21E,23E,25E,27S,28R)-4,6,8,10,12,14,16,27-octahydroxy-3-((R)-1-hydroxyhexyl)-17,28-dimethyloxacyclooctacosa-17,19,21,23,25-pentaen-2-one | ||
| Canonical SMILES | O[C@@H]1/C(C)=C/C=C/C=C/C=C/C=C/[C@H](O)[C@@H](C)OC([C@@]([C@H](O)CCCCC)([H])[C@@H](O)C[C@@H](O)C[C@@H](O)C[C@@H](O)C[C@@H](O)C[C@@H](O)C1)=O | ||
| 分子式 | C35H58O11 | 分子量 | 654.83 |
| 溶解度 | 1mg/ml in Ethanol; 5mg/ml in DMF; 10mg/ml in DMSO | 储存条件 | Store at -20°C, protect from light. We recommend using the stock solution within 24 hours or it may result in reduced activity. |
| 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 | 1.5271 mL | 7.6356 mL | 15.2711 mL |
| 5 mM | 0.3054 mL | 1.5271 mL | 3.0542 mL |
| 10 mM | 0.1527 mL | 0.7636 mL | 1.5271 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。