BODIPY 558/568 C₁₂；Red C₁₂; BODIPY 493/503；Nile Red 尼罗红；Neutral lipds 中性脂滴；DiI 细胞膜探针；DiR； CAS NO：158757-84-7

产品信息

产品名称		规格	CAS NO.
BODIPY 558/568 C12 脂滴荧光探针		1mg	158757-84-7

产品描述

BODIPY 558/568 C₁₂是一种偶联脂肪酸的荧光探针，用于脂滴检测，最大激发和发射波长分别为558/568nm，适用于监测活细胞内的脂滴定位和动力学。

产品特性

1）CAS NO.：158757-84-7

2）化学名：Borate(1-), difluoro[5-[[5-(2-thienyl)-2H-pyrrol-2-ylidene-[kappa]N]methyl]-1H-pyrrole-2-dodecanoato(2-)-[kappa]N1]-, hydrogen,(T-4)-

3）同义名：Red C₁₂

4）分子式：C₂₅H₃₁BF₂N₂O₂S

5）分子量：472.4

6）纯度：≥95%

7）Ex/Em：558/568nm

8）外观：油状

9）溶解性：溶于DMSO或DMF

9）化学结构式：

保存与运输方法

保存：-20ºC避光干燥保存，至少2年有效。 

运输：冰袋运输。

储存液的制备和保存

将低温保存的BODIPY 558/568 C₁₂（Mw: 472.4）置于室温回温至少20min，低速离心后加入一定量的无水DMSO配制成适量浓度的母液，比如5mM（往1mg 粉末加入423.37μl DMSO，充分溶解即可）。根据单次用量将储存液分装，≤-20℃避光干燥保存。需注意溶液内湿度的逐渐积累会随着时间推移引起探针聚集，因而务必干燥保存储存液。

探针的标记（仅作参考）

1. 由于BODIPY 558/568 C₁₂属于疏水染料，难以快速的分散进入水溶性溶液中，为了能均匀稳定的标记细胞，可参考以下方法标记活细胞。
2. 2.探针的工作浓度建议是1-10µM，加载时间根据实际的应用来调整，可查阅相关文献资料。

应用示例（来自文献，仅作参考）

1. 文献来源：Qi G, Mi Y, Shi X, Gu H, Brinton RD, Yin F. ApoE4 Impairs Neuron-Astrocyte Coupling of Fatty Acid Metabolism. Cell Rep. 2021;34(1):108572. doi:10.1016/j.celrep.2020.108572

使用目的：分析细胞间的脂肪酸转移。

使用方法：DIV7 ApoE3 或ApoE4 神经元或成年星形胶质细胞（0.5×10⁶ cells/well，6孔板）用含2μM BODIPY 558/568 C₁₂的培养基孵育16h，之后用PBS清洗3次。这些标记的细胞（供体）再转移到先前培养的未标记神经元或成年星形胶质细胞（受体，10⁵ cells，22mm爬片）4h。爬片上的细胞用PBS清洗后再做固定。荧光显微镜成像分析。

2.文献来源：Ioannou MS, Liu Z, Lippincott-Schwartz J. A Neuron-Glia Co-culture System for Studying Intercellular Lipid Transport. Curr Protoc Cell Biol. 2019 Sep;84(1):e95. doi: 10.1002/cpcb.95. PMID: 31483110.

使用目的：分析神经元到胶质细胞的脂质转移。

使用方法：实验前一天，用含2.5μM BODIPY 558/568 C₁₂的神经元培养基加载神经元；18h之后用预热的DPBS清洗神经元2次，再用新鲜培养基37℃孵育1h；分别用DPBS清洗生长在盖玻片上的神经元和胶质细胞；加预热的HBSS到胶质细胞，使用无菌镊子夹住长有神经元的盖玻片，盖到长有星形胶质细胞的盖玻片上；37℃孵育这一夹心培养物4h；用镊子轻轻夹走神经元盖玻片；为了固定胶质细胞，先用DPBS清洗2遍，用3%多聚甲醛固定10min，用DPBS清洗2次；用含5µg/ml BODIPY 493/503的DPBS孵育固定的胶质细胞，室温避光孵育10min，用DPBS清洗2次；

Fig. Schematic of fatty acid transfer assay. Neurons are incubated with Red-C12 overnight and then incubated with glia on separate coverslips for 4 hr. Glia are fixed, and the appearance of Red-C12 in astrocytic lipid droplets is imaged and quantified. This figure is reproduced from Ioannou et al. (2019).

Fig. Appearance of neuron-derived fatty acids (Red-C12) in glial lipid droplets. After the transfer assay, glia were fixed, stained with BODIPY 493/503 (BD-493) to label lipid droplets, and imaged using a Zeiss 880 confocal microscope with a 63× objective.

文献来源：Rambold AS, Cohen S, Lippincott-Schwartz J. Fatty acid trafficking in starved cells: regulation by lipid droplet lipolysis, autophagy, and mitochondrial fusion dynamics. Dev Cell. 2015 Mar 23;32(6):678-92. doi: 10.1016/j.devcel.2015.01.029. Epub 2015 Mar 5. PMID: 25752962; 

使用目的：脂肪酸脉冲追踪和共培养实验（Fluorescent FA pulse-chase and co-culture experiments）。

使用方法：MEFs were incubated with complete medium (DMEM with 10% FBS and 4 mM glutamine, CM) containing 1 μM BODIPY 558/568 C₁₂ (Red C12) or 2 μM β-BODIPY FL C₁₂-HPC (FL HPC) for 16 h. Cells were then washed three times with CM, incubated for 1 h in order to allow the fluorescent lipids to incorporate into LDs or cellular membranes, and then chased for the time indicated in CM or Hank’s buffered saline solution (HBSS) in the absence or presence of various drugs. Mitochondria were labeled with 100 nM MitoTracker Green FM for 30 minutes prior to imaging. To label LDs, BODIPY 493/503 or BODIPY665/676 was added to cells at 200 ng/ml immediately prior to imaging and was present during imaging. 

Fig. Fatty acid trafficking can be visualized using a fluorescent fatty acid pulse-chase assay

(A) Schematic representation of the fluorescent FA pulse-chase assay: cells were pulsed with Red C12 overnight, washed, and incubated with CM for 1 h in order to allow the Red C12 to accumulate in LDs. Cells were then chased in CM or HBSS for the indicated periods of time and imaged to determine the subcellular localization of the FA. (B–D) WT MEFs were assayed as described in Figure 1A and chased in CM or HBSS for 0 h, 6 h, or 24 h. LDs were labeled using (B) BODIPY 493/503 and (C) mitochondria were labeled using MitoTracker Green. Scale bar, 10μm. (D) Relative cellular localization of Red C12 was quantified by Pearson’s coefficient analysis. Data were expressed as means ± SEM. (E) TLC resolving Red C12 isolated from WT MEFs assayed as described above and chased for 6 h or 24 h with HBSS in the absence or presence of etomoxir, see also Figure S1.

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。

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