耗材描述
ER-Tracker Blue-White DPX
产品活性:ER-Tracker 染料是与 Glibenclamide (HY-15206) 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。
产品来源: https://www.medchemexpress.cn/er-tracker-blue-white-dpx.html
研究领域:Others
作用靶点:Fluorescent Dye
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 1×106/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.
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