ER-Tracker 染料是与 Glibenclamide 偶联的 BODIPY 列染料的衍生物，对内质网高选择性结合，在低浓度下对细胞无毒性，该类染料属于环境敏感型的探针，与甲醛处理后仍能保留部分荧光，具有荧光寿命高，消光系数好等特点。Glibenclamide 是一种 ATP 依赖性 K+ 通道阻滞剂 (Kir6, KATP) 和 CFTR Cl-通道阻滞剂，在内质网中发生结合。ER-Tracker 不适合对固定后细胞进行染色。 ER-Tracker dye is a derivative of BODIPY series dyes coupled with Glibenclamide , 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. In Vitro 1. Preparation of ER-Tracker working solution 1.1 Preparation of the stock solution Dissolve 100 ug ER-Tracker in 128 μL DMSO to obtain 1 mM of stock solution. Note: It is recommended to store the stock solution at -20℃or -80℃away from light and avoid repetitive freeze-thaw cycles. 1.2 Preparation of ER-Tracker working solution Dilute the stock solution in serum-free cell culture medium or PBS to obtain 100 nM-1 μM of working solution. Note: Please adjust the concentration of ER-Tracker working solution according to the actual situation. 2. Cell staining 2.1 Suspension cells (6-well plate) a. Centrifuge at 1000 g at 4℃for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.The cell density is 1×10 6 /mL. b. Add 1 mL of working solution, and then incubate at room temperature for 5-30 minutes. c. Centrifuge at 400 g at 4℃for 3-4 minutes and then discard the supernatant. d. Wash twice with PBS, 5 minutes each time. e. Resuspend cells with serum-free cell culture medium or PBS. Observation by fluorescence microscopy or flow cytometry. 2.2 Adherent cells a. Culture adherent cells on sterile coverslips. b. Remove the coverslip from the medium and aspirate excess medium. c. Add 100 μL of working solution, gently shake it to completely cover the cells,and then incubate at room temperature for 5-30 minutes. d. Wash twice with medium, 5 minutes each time. Observation by fluorescence microscopy or flow cytometry. Note: If detection by flow cytometry, cells need to be resuspended before staining. MCE has not independently confirmed the accuracy of these methods. They are for reference only. Form:Solid