Hydroxyphenyl Fluorescein  (Synonyms: HPF; 3'-p-(Hydroxyphenyl) fluorescein)

Hydroxyphenyl Fluorescein (HPF) is a stable ROS fluorescent probe dye. Hydroxyphenyl Fluorescein has stronger specificity and stability than H2DCFDA (HY-D0940). Hydroxyphenyl Fluorescein can produce strong green fluorescence through hydroxyl radical reaction with intracellular peroxynitroso. Hydroxyphenyl Fluorescein can be applied for fluorescence microscopy, high-throughput imager, luciferase microplate reader or flow cytometry. Ex/Em=490/515 nm^[1].

HPF 工作液的配制
1.1 制备储存液
用 DMSO 配制 10 mM 的 HPF。
注：HPF 储存液建议分装后于 -20 ℃ 或 -80 ℃ 避光保存。
1.2 工作液的配制
用预热好的无血清细胞培养基或 PBS 稀释储存液，配制成 1-10 μM 的 HPF 工作液。
注：请根据实际情况调整 HPF 工作液浓度，且现用现配。

细胞染色
2.1 准备工作
悬浮细胞：离心收集细胞，加入 PBS 洗涤两次，每次 5 分钟。
贴壁细胞：弃去培养基，加入胰蛋白酶消化细胞。离心弃去上清后，加入 PBS 洗涤两次，每次 5 分钟。
2.2 加入 1 mL HPF 工作液，室温孵育 15 分钟。
2.3 400 g，4℃ 离心 3-4 分钟，弃去上清。
2.4 加入 PBS 洗涤细胞两次，每次 5 分钟。
2.5 用 1 mL 无血清培养基或 PBS 重悬细胞后，在荧光显微镜或流式细胞仪下检测。

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

The Hydroxyphenyl Fluorescein (HPF) probe (15 μl, 25 μM) is given intratesticularly in anaesthetized mice 20 min before IR (ionizing radiation). The accumulation of OH by the fluorescence signal emitted by the oxidized form of HPF is assessed^[3].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

424.40

C₂₆H₁₆O₆

359010-69-8

液体

Colorless to light yellow

515

490

羟苯基荧光素

Room temperature in continental US; may vary elsewhere.

Solution, -20°C, protect from light, 2 years

• [1]. Ming-De Li, et al. Dynamics of Oxygen-Independent Photocleavage of Blebbistatin as a One-Photon Blue or Two-Photon Near-Infrared Light-Gated Hydroxyl Radical Photocage. J Am Chem Soc. 2018 Nov 21;140(46):15957-15968.  [Content Brief]