1,3-Diphenylisobenzofuran (solution) 

1,3-Diphenylisobenzofuran (DPBF) (solution) has been developed as a selective probe for the detection and quantitative determination of hydrogen peroxide in samples containing different reactive nitrogen and oxygen species (RNOS). DPBF is a fluorescent probe which, for almost 20 years, was believed to react in a highly specific manner toward some reactive oxygen species such as singlet oxygen and hydroxy, alkyloxy or alkylperoxy radicals^[1].
Solvent and concentration: DMSO: 10 mM
The 1 mL volume is defined as the base specification. All larger sizes correspond to incremental volumes of this base.

使用方法
1,3-Diphenylisobenzofuran (DPBF) 工作液的配制
1.1 制备储存液
注：DPBF 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 DPBF 工作液浓度，且现用现配。

细胞染色
2.1 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。
注：如不做流式实验，贴壁细胞可不做消化处理。
2.2 加入 1 mL DPBF 工作液，室温孵育 30 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。

注意事项
1. DPBF 储存液建议分装后于 -20°C 或 -80°C 避光保存，避免反复冻融。
2. 请根据实际情况调整 DPBF 工作液浓度。
3. 本产品仅限于专业人员的科学研究用，不得用于临床诊断或治疗，不得用于食品或药品。
4. 为了您的安全和健康，请穿实验服并戴一次性手套操作。

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

270.33

C₂₀H₁₄O

5471-63-6

Room temperature in continental US; may vary elsewhere.

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

• [1]. Weili Fan, et al. Calcium carbonate-methylene blue nanohybrids for photodynamic therapy and ultrasound imaging. Sci China Life Sci. 2018 Apr;61(4):483-491.  [Content Brief]

  [2]. P. Carloni, et al. On the use of 1,3-diphenylisobenzofuran (DPBF). Reactions with carbon and oxygen centered radicals in model and natural systems. Research on Chemical Intermediates volume 19, pages395-405(1993).

  [3]. Bai X, et al. HKOH-1: A Highly Sensitive and Selective Fluorescent Probe for Detecting Endogenous Hydroxyl Radicals in Living Cells. Angew Chem Int Ed Engl. 2017 Oct 9;56(42):12873-12877.  [Content Brief]