Dihydroethidium (solution) 

Dihydroethidium (solution) , also known as DHE, is a peroxide indicator. Dihydroethidium penetrates cell membranes to form a fluorescent protein complex with blue fluoresces. After entering the cells, Dihydroethidium is mainly localized in the cell membrane, cytoplasm and nucleus, and the staining effect is the strongest in the nucleus. Dihydroethidium produces inherent blue fluorescence with a maximum excitation wavelength of 370 nm and a maximum emission wavelength of 420 nm; after dehydrogenation, Dihydroethidium combines with RNA or DNA to produce red fluorescence with a maximum excitation wavelength of 300 nm and a maximum emission wavelength of 610 nm. 535 nm can also be used as the excitation wavelength for actual observation^[1].
Solvent and concentration: DMSO: 5 mM

一般方案
1.1 Dihydroethidium 工作液的制备
稀释在无血清细胞培养基或 PBS 中加入 1-10 μM 的 Dihydroethidium 工作液。
注意：请根据实际情况调整 Dihydroethidium 工作液的浓度。
细胞染色
2.1 悬浮细胞：4°C，1000g 离心 3-5 分钟，弃上清。PBS 洗涤 2 次，每次 5 分钟。
贴壁细胞：弃去细胞培养基，加入胰蛋白酶解离细胞，制成单细胞悬液。在 4°C 下以 1000 g 离心 3-5 分钟，然后弃去上清液。PBS 洗涤 2 次，每次 5 分钟。
2.2 加入 1mL Dihydroethidium 工作液，室温孵育 30 分钟。
2.3 4°C，400g 离心 3-4 分钟，弃上清。
2.4 PBS 洗涤 2 次，每次 5 分钟。
2.5 用无血清细胞培养基或 PBS 重悬细胞，荧光显微镜或流式细胞仪检测。

储存
-20°C，1 年
避光

注意事项
1. 建议将原液储存于 -20°C 或 -80°C 避光，避免反复冻融。
2.请根据实际情况调整 Dihydroethidium 工作液的浓度。
3.本品供研发使用
4. 为了您的安全和健康，请穿着实验服和一次性手套操作。

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

315.42

C₂₁H₂₁N₃

104821-25-2

Room temperature in continental US; may vary elsewhere.

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

• [1]. Zielonka J, et al. Global profiling of reactive oxygen and nitrogen species in biological systems: high-throughput real-time analyses. J Biol Chem. 2012 Jan 27;287(5):2984-95.  [Content Brief]