Indo-1 AM  (Synonyms: Indo-1 Acetoxymethyl ester)

Calcium is an important part of the human body, usually in the form of calcium, a large number of bones and teeth of the human body, a small amount of blood and tissues. MCE calcium ion detection probe can specifically identify intracellular calcium ions, with high sensitivity, low cytotoxicity, increased AM acetylmethyl ester can enter the cell well, after being sheared by the intracellular esterase stay in the cell to bind to calcium ions, produce strong fluorescence, in addition, Mag-indo-1/AM and Mag-Fluo-4 AM at a certain concentration (usually 5 mM) can effectively identify intracellular magnesium ions^[1].

1. Indo-1 AM 工作液的配制
1.1 制备储存液
用无水 DMSO 稀释 Indo-1 AM 去配制 10 mM 的储备液。
注：Indo-1 AM 储存液建议分装后于 -20 ℃ 或 -80 ℃ 避光保存。
1.2 工作液的配制
用预热好的 HBSS 稀释储存液，配制成 1-10 μM 的 Indo-1 AM 工作液。
注：请根据实际情况调整 Indo-1 AM 工作液浓度，且现用现配。
2. 细胞染色(悬浮细胞)
2.1 离心收集细胞，加入 PBS 洗涤两次，每次 5 分钟。细胞密度在 1×10⁶/mL。
2.2 加入 1 mL Indo-1 AM 工作液，室温孵育 5-30 分钟。
2.3 400 g，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 HBSS 重悬细胞后，使用荧光显微镜或流式细胞仪进行观察。
3. 细胞染色(贴壁细胞)
3.1 将贴壁细胞培养于无菌盖玻片上。
3.2 从培养基中移出盖玻片，吸除多余培养基。
3.3 加入 100 μL 染料工作液，轻轻晃动使其完全覆盖细胞，孵育 5-30 分钟。
3.4 吸去染料工作液，用培养基洗 2-3 次，每次 5 分钟，使用荧光显微镜进行观察。

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

1009.91

C₄₇H₅₁N₃O₂₂

112926-02-0

固体

Light yellow to yellow

410

364

Room temperature in continental US; may vary elsewhere.

-20°C, protect from light

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

• [1]. Andrienko T, et al. Real-Time Fluorescence Measurements of ROS and [Ca2+] in Ischemic / Reperfused Rat Hearts: Detectable Increases Occur only after Mitochondrial Pore Opening and Are Attenuated by Ischemic Preconditioning. PLoS One. 2016 Dec 1;11(12):e01  [Content Brief]

  [2]. Andrienko T, et al. Real-Time Fluorescence Measurements of ROS and [Ca2+] in Ischemic / Reperfused Rat Hearts: Detectable Increases Occur only after Mitochondrial Pore Opening and Are Attenuated by Ischemic Preconditioning. PLoS One. 2016 Dec 1;11(12):e0167300.  [Content Brief]

  [3]. Blatter LA, et al. Intracellular diffusion, binding, and compartmentalization of the fluorescent calcium indicators indo-1 and fura-2. Biophys J. 1990 Dec;58(6):1491-9.  [Content Brief]