DAF-FM DA (solution) 

DAF-FM DA (Diaminofluorescein-FM diacetate) (solution) is a fluorescent probe for the detection and bioimaging of nitric oxide (NO). DAF-FM DA spontaneously crosses the plasma membrane and is subsequently cleaved by esterases to generate intracellular DAF-FM (Ex/Em=495/515 nm) ^[1].
Solvent and concentration: DMSO: 2 mM

指南 (以下是我们推荐的方案，本方案仅提供指南，应根据您的具体需要进行修改)。
1.1 储存液^[2]
注: DAF-FM DA 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 Hank’s 缓冲液或无血清培养基稀释储存液，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 DAF-FM DA 工作液浓度，且现用现配。
2 染色步骤
1) 将乳腺癌细胞接种于 6 孔板中。
2) 吸除培养基。
3) 每孔加入 5 μM 的 DAF-FM DA 工作液，于 37°C 黑暗中孵育 30 min，使探针进入细胞。
4) 更换为新鲜的 Hank’s 缓冲液，继续孵育 20 min，完成去酯化反应。
5) Hoechst 染料对细胞核进行复染。
6) 使用酶标仪在 488 nm 激发光和 515 nm 发射光下定量荧光强度，以 Hoechst 染色的细胞数进行归一化。

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

使用经 DAF-FM DA (5 μM) 活染色的斑马鱼胚胎切片，可以确认荧光信号主要位于正在进行的骨形成区域^[1]。

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

496.42

C₂₅H₁₈F₂N₂O₇

254109-22-3

Room temperature in continental US; may vary elsewhere.

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

• [1]. Zhi Xu, et al. RelB-activated GPX4 inhibits ferroptosis and confers tamoxifen resistance in breast cancer. Redox Biol. 2023 Dec:68:102952.  [Content Brief]

  [2]. Ann Huysseune, et al. Bone Formation in Zebrafish: The Significance of DAF-FM DA Staining for Nitric Oxide Detection. Biomolecules. 2023 Dec 12;13(12):1780.  [Content Brief]