摘要：ABDP 493/503 (同BODIPY 493/503) 是一种亲脂性荧光探针，可以用作中性脂质的染色剂，以及油和其他非极性脂质的示踪剂。最大激发和发射波长分别是493/503nm，适用于活细胞和固定细胞标记。
ABDP 493/503 (同BODIPY 493/503) 是一种亲脂性荧光探针，可以用作中性脂质的染色剂，以及油和其他非极性脂质的示踪剂。最大激发和发射波长分别是493/503nm，适用于活细胞和固定细胞标记。
一、生物活性
ABDP 493/503是一种新型的、性能优异的中性脂滴特异性荧光探针。它在设计上属于“开启式”探针，其核心机理在于其分子结构在自由状态下会发生分子内运动，导致荧光被猝灭；而当其进入疏水性的脂滴内部环境中时，分子运动受到限制，从而恢复强烈的荧光信号。这种特性使其具有极低的背景荧光和极高的信噪比，非常适合用于高分辨率成像和活细胞动态追踪。ABDP 493/503的命名来源于其光谱特性：其最大激发波长在493纳米左右，最大发射波长在503纳米左右，恰好与常用的FITC滤光片组相匹配，这使得它可以在大多数常规激光共聚焦显微镜上方便地使用。与传统广泛使用的脂滴染料如BODIPY 493/503相比，ABDP 493/503展现出更优异的光稳定性和更高的脂滴特异性，能更清晰、更持久地标记细胞内的中性脂滴，而不会像一些染料那样容易浸出或发生非特异性染色。因此，它已成为研究脂肪代谢、肥胖、肝steatosis、以及评估细胞脂质储存与分解等生命科学和医学研究领域的强大工具，为科研人员提供了更可靠、更灵敏的成像解决方案。
二、化学性质/溶解性/储存
分子量：262.11
分子式： C14H17BF2N2
CAS号： 121207-31-6
中文名称： 中性脂滴荧光探针
储存条件 ：-20°C, protect from light, dry, sealed
运输方式： 冰袋运输，根据产品的不同，可能会有相应调整。
*不同实验中用到的溶剂可能不同，具体实验所需溶剂及溶解方法请参考相关文献描述。
三、储存液的制备和保存
1）将低温保存的 ABDP 493/503 置于室温回温约20min，低速离心后加入一定量的无水乙醇或无水DMSO配制成适量浓度的母液，比如5mM（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引起探针聚集，从而务必干燥保存储存液。
2）如需长期保存，可以用无水乙醇溶解粉末，之后分装到小量，使用真空泵来挥发掉乙醇。置于≤-20℃避光干燥保存。
BODIPY染色用于流式细胞仪（摘自公开文献，仅供参考）
1. Grow cells under culture conditions relevant for the study. A 35 mm dish/well is sufficient for the cell numbers required in this assay. For our assays, 50,000 A498 cells in 35 mm well were sufficient. Overnight incubation of cells with 30 μM oleic acid can serve as a positive control for increased neutral lipid content, as oleic acid is a potent inducer of triglyceride synthesis and storage. Fatty acid free BSA serves as a control.
2. At the time-point of interest, prepare 2 μM BODIPY staining solution in PBS. The volume of staining solution required for each sample corresponds to the volume of media used for culturing cells.
3. Wash cells with a quick rinse using 3 ml PBS to remove media/serum.
4. Incubate on BODIPY staining solution in the dark for 15 min at 37 °C. Include an unstained control for flow cytometry.
Note: From this point, protect samples from light as much as possible.
5. Wash cells with a quick rinse using 3 ml PBS to remove staining solution.
6. Trypsinize cells to generate a single cell suspension. For the A498 cell line used in this protocol, cells were incubated with Trypsin-EDTA (0.25%) for 5 min at 37 °C.
7. Add 5 ml of PBS and transfer cell suspension to a 15 ml conical tube.
8. Pellet cells at 250 × g, 5 min, 4 °C.
9. Aspirate supernatant, wash the cell pellet with a quick rinse using 3 ml PBS, and pellet cells at 250 × g, 5 min, 4 °C.
10. Carefully aspirate the supernatant and resuspend cells in 300 μl 1× flow cytometry buffer.
11. Pass cell suspension through a 35 μm filter into a FACS tube.
12. Perform flow cytometry. Obtain a minimum of 10,000 events per condition.
13. The investigator can analyze data as mean fluorescence or display the data as a histogram.
来源：AbMole