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].

指南 (以下為我們推薦的實(shí)驗(yàn)方案。本方案僅作為參考指南，具體操作應(yīng)根據(jù)您的實(shí)際需求進(jìn)行調(diào)整)。
1. Indo-1 AM 工作液的配制
1.1 制備儲(chǔ)存液
用無(wú)水 DMSO 稀釋 Indo-1 AM 去配制 10 mM 的儲(chǔ)備液。
1.2 工作液的配制
用預(yù)熱好的 HBSS 稀釋儲(chǔ)存液，配制成 1-10 μM 的 Indo-1 AM 工作液。
注：請(qǐng)根據(jù)實(shí)際情況調(diào)整 Indo-1 AM 工作液濃度，且現(xiàn)用現(xiàn)配。
2. 細(xì)胞染色 (懸浮細(xì)胞)
2.1 在 4°C，1000 g 條件下離心 3-5 分鐘，收集細(xì)胞沉淀，加入 PBS 洗滌兩次，每次 5 分鐘。細(xì)胞密度在 1×10⁶/mL。
2.2 加入 1 mL Indo-1 AM 工作液，室溫孵育 5-30 分鐘。
2.3 在 4°C，400 g 條件下，離心 3-4 分鐘，棄去上清。
2.4 加入 PBS 洗滌細(xì)胞兩次，每次 5 分鐘。
2.5 用 1 mL HBSS 重懸細(xì)胞后，使用熒光顯微鏡或流式細(xì)胞儀進(jìn)行觀察。
3. 細(xì)胞染色 (貼壁細(xì)胞)
3.1 將貼壁細(xì)胞培養(yǎng)于無(wú)菌蓋玻片上。
3.2 從培養(yǎng)基中移出蓋玻片，吸除多余培養(yǎng)基。
3.3 加入 100 μL 染料工作液，輕輕晃動(dòng)使其完全覆蓋細(xì)胞，孵育 5-30 分鐘。
3.4 吸去染料工作液，用培養(yǎng)基洗 2-3 次，每次 5 分鐘，使用熒光顯微鏡進(jìn)行觀察。

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

1009.91

C₄₇H₅₁N₃O₂₂

112926-02-0

固體

Light yellow to yellow

410

364

Room temperature in continental US; may vary elsewhere.

• [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]