BODIPY 558/568 C₁₂ 脂滴熒光探針

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產品標簽

BODIPY 558/568 C₁₂；Red C₁₂; BODIPY 493/503；Nile Red 尼羅紅；Neutral lipds 中性脂滴；DiI 細胞膜探針；DiR； CAS NO：158757-84-7

產品信息

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產品描述

BODIPY 558/568 C₁₂是一種偶聯脂肪酸的熒光探針，用于脂滴檢測，最大激發和發射波長分別為558/568nm，適用于監測活細胞內的脂滴定位和動力學。

產品特性

1）CAS NO.：158757-84-7

2）化學名：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）同義名：Red C₁₂

4）分子式：C₂₅H₃₁BF₂N₂O₂S

5）分子量：472.4

6）純度：≥95%

7）Ex/Em：558/568nm

8）外觀：油狀

9）溶解性：溶于DMSO或DMF

9）化學結構式：

保存與運輸方法

保存：-20oC避光干燥保存，至少2年有效。 

運輸：冰袋運輸。

儲存液的制備和保存

將低溫保存的BODIPY 558/568 C₁₂（Mw: 472.4）置于室溫回溫至少20min，低速離心后加入一定量的無水DMSO配制成適量濃度的母液，比如5mM（往1mg 粉末加入423.37μl DMSO，充分溶解即可）。根據單次用量將儲存液分裝，≤-20℃避光干燥保存。需注意溶液內濕度的逐漸積累會隨著時間推移引起探針聚集，因而務必干燥保存儲存液。

探針的標記（僅作參考）

1. 由于BODIPY 558/568 C₁₂屬于疏水染料，難以快速的分散進入水溶性溶液中，為了能均勻穩定的標記細胞，可參考以下方法標記活細胞。

2. 探針的工作濃度建議是1-10µM，加載時間根據實際的應用來調整，可查閱相關文獻資料。

應用示例（來自文獻，僅作參考）

1. 文獻來源：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：分析細胞間的脂肪酸轉移。

使用方法：DIV7 ApoE3 或ApoE4 神經元或成年星形膠質細胞（0.5×10⁶ cells/well，6孔板）用含2μM BODIPY 558/568 C₁₂的培養基孵育16h，之后用PBS清洗3次。這些標記的細胞（供體）再轉移到先前培養的未標記神經元或成年星形膠質細胞（受體，10⁵ cells，22mm爬片）4h。爬片上的細胞用PBS清洗后再做固定。熒光顯微鏡成像分析。

2.文獻來源：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：分析神經元到膠質細胞的脂質轉移。

使用方法：實驗前一天，用含2.5μM BODIPY 558/568 C₁₂的神經元培養基加載神經元；18h之后用預熱的DPBS清洗神經元2次，再用新鮮培養基37℃孵育1h；分別用DPBS清洗生長在蓋玻片上的神經元和膠質細胞；加預熱的HBSS到膠質細胞，使用無菌鑷子夾住長有神經元的蓋玻片，蓋到長有星形膠質細胞的蓋玻片上；37℃孵育這一夾心培養物4h；用鑷子輕輕夾走神經元蓋玻片；為了固定膠質細胞，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孵育固定的膠質細胞，室溫避光孵育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).maokangbio

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.

3. 文獻來源：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：脂肪酸脈沖追蹤和共培養實驗（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. maokangbio

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.

注意事項

1）熒光染料均存在淬滅問題，請盡量注意避光，以減緩熒光淬滅。

2）為了您的安全和健康，請穿實驗服并戴一次性手套操作。

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