Hydroxyphenyl Fluorescein (solution) 

Hydroxyphenyl Fluorescein (HPF) (solution) 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].
Solvent and concentration: DMSO: 10 mM

HPF 工作液的配制
1.1 保存储存液
HPF 储存液建议分装后于 -20 °C 或 -80 °C 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，可根据实际情况稀释相应的储备液。注意，如果溶剂为 DMSO，则必须考虑 DMSO 的细胞毒性，并应制备溶剂对照；如果溶剂为纯水，则工作液在加入细胞前需要过滤并除菌。
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°C 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.

424.41

C₂₆H₁₆O₆

359010-69-8

Room temperature in continental US; may vary elsewhere.

Please store the product under the recommended conditions in the Certificate of Analysis.

• [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]