ER-Tracker Red (solution) 

ER-Tracker Red (solution) is a derivative of BODIPY series dyes coupled with Glibenclamide (HY-15206), highly selective binding to the endoplasmic reticulum, non-toxic to cells at low concentrations, this type of dye is an environmentally sensitive probe, and formaldehyde treatment can still retain part of the fluorescence, with high fluorescence life, good extinction coefficient and other characteristics. Glibenclamide is an atp-dependent K⁺ channel blocker (Kir6, KATP) and CFTR Cl-channel blocker that binds in the endoplasmic reticulum. ER-Tracker is not suitable for staining cells after fixation. Ex/Em=587/615 nm^[1].

1.ER-Tracker 工作液的配制
1.1 制备储存液
用 109 μL 无水 DMSO 稀释 100 μg ER-Tracker 去配制 1 mM 的储备液。
注：ER-Tracker 储存液建议分装后于 -20℃ 或 -80℃ 避光保存。
1.2 工作液的配制
用预热好的无血清细胞培养基或 PBS 稀释储存液，配制成 100 nM-1 μM 的 ER-Tracker工作液。
注：请根据实际情况调整 ER-Tracker 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 ER-Tracker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 无血清培养基或 PBS 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。
注：若需要用流式细胞仪检测，需将细胞用胰蛋白酶消化重悬后再进行染色。

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

915.23

C₄₄H₄₂BClF₂N₆O₇S₂

Room temperature in continental US; may vary elsewhere.

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

• [1]. Merianda TT, et al. A functional equivalent of endoplasmic reticulum and Golgi in axons for secretion of locally synthesized proteins. Mol Cell Neurosci. 2009 Feb;40(2):128-42.

  [2]. Adiki Raja Sekhar, et al. A cell-permeant small molecule for the super-resolution imaging of the endoplasmic reticulum in live cells. Org Biomol Chem. 2019 Apr 10;17(15):3732-3736.  [Content Brief]

  [3]. Tanuja T Merianda, et al. A functional equivalent of endoplasmic reticulum and Golgi in axons for secretion of locally synthesized proteins. Mol Cell Neurosci. 2009 Feb;40(2):128-42.  [Content Brief]