MitoSOX Red (solution) 

MitoSOX Red (solution) is a live cell fluorescent probe that specifically targets mitochondria and is cell membrane permeable. MitoSOX Red enters mitochondria and is oxidized by superoxide but not by other ROS or RNS generating systems. The oxidized MitoSOX Red then binds to nucleic acids in mitochondria/nucleus, producing strong red fluorescence. MitoSOX Red can be used as a fluorescent indicator to specifically detect superoxide. In addition, superoxide dismutase (SOD) can prevent the oxidation of MitoSOX Red.
Excitation/emission wavelength: 510/580 nm.
Solvent and concentration: DMSO: 1 mM.

实验方案

1. MitoSOX Red 工作液的制备
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。
2. 细胞染色
2.1 悬浮细胞 (6 孔板)
a．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。细胞密度为 1×10⁶/mL。
b．加入 1 mL 工作液，室温孵育 5-30 分钟。
c． 4°C、400 g 离心 3-4 分钟，弃上清。
d．用 PBS 洗涤 2 次，每次5分钟。
e．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术观察。
2.2 贴壁细胞
a．在无菌盖玻片上培养贴壁细胞。
b.从培养基上取下盖玻片并吸出多余的培养基。
c.加入 100 μL 工作液，轻摇使之完全覆盖细胞，室温孵育 5-30 分钟。
d．用培养基洗两次，每次 5 分钟。荧光显微镜或流式细胞术观察。

储存
分装保存，避免反复冻融，-20°C，1 年。避光

注意事项
1、请根据实际情况调整 MitoSOX Red 工作液的浓度。
2、本品仅供 R&D 使用，不得用于药物、家庭或其他用途。
3. 为了您的安全和健康，请穿白大褂，戴一次性手套操作。

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

Room temperature in continental US; may vary elsewhere.

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

• [1]. Price OT, et al. Quantitative fluorescence of 5-FU-treated fetal rat limbs using confocal laser scanning microscopy and Lysotracker Red. Cytometry A. 2003;53(1):9-21.  [Content Brief]

  [2]. Fengjiao Jin, et al. The PI3K/Akt/GSK-3β/ROS/eIF2B pathway promotes breast cancer growth and metastasis via suppression of NK cell cytotoxicity and tumor cell susceptibility. Cancer Biol Med. 2019 Feb;16(1):38-54.  [Content Brief]