ER-Tracker Blue-White DPX 

ER-Tracker dye 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 fixed cells. Staining followed by fixation is possible, but cells fixed with aldehydes will only retain partial fluorescence (Ex/Em = 374/ 430-640 nm)^[1].

Guide (The following is our recommended protocol. This protocol only provides guidance and should be modified according to your specific needs).
1. Preparation of ER-Tracker working solution
1.1 Preparation of stock solution
Dissolve 100 μg ER-Tracker in 172 μL anhydrous DMSO to prepare a 1 mM stock solution.
Note: It is recommended to aliquot the ER-Tracker stock solution and store at -20°C or -80°C, protected from light.
1.2 Preparation of working solution
Dilute the stock solution with pre-warmed serum-free cell culture medium or PBS to prepare a 100 nM-1 μM ER-Tracker working solution.
Note: Please adjust the concentration of the ER-Tracker working solution according to actual conditions and prepare fresh before use.
2. Cell staining (suspension cells)
2.1 Collect cells by centrifugation, wash twice with PBS for 5 minutes each time. Adjust the cell density to 1×10⁶/mL.
2.2 Add 1 mL ER-Tracker working solution and incubate at room temperature for 5-30 minutes.
2.3 Centrifuge at 400 g for 3-4 minutes and discard the supernatant.
2.4 Wash the cells twice with PBS for 5 minutes each time.
2.5 Resuspend the cells in 1 mL serum-free medium or PBS, then observe using a fluorescence microscope or flow cytometer.
3. Cell staining (adherent cells)
3.1 Culture adherent cells on sterile coverslips.
3.2 Remove the coverslips from the culture medium and aspirate excess medium.
3.3 Add 100 μL staining working solution, gently shake to ensure complete coverage of the cells, and incubate for 5-30 minutes.
3.4 Remove the staining working solution and wash with culture medium 2-3 times for 5 minutes each time, then observe using a fluorescence microscope.
Note: If flow cytometry analysis is required, digest the cells with trypsin, resuspend, and then proceed with staining.

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

580.53

C₂₆H₂₁F₅N₄O₄S

485

430

287715-95-1

Solid

Light yellow to yellow

O=S(C(C=C1)=CC=C1C(O2)=NC=C2C3=CC=C(N(C)C)C=C3)(NCCNC(C4=C(F)C(F)=C(F)C(F)=C4F)=O)=O

Room temperature in continental US; may vary elsewhere.

4°C, sealed storage, away from moisture and light

*In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

• [1]. 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]

  [2]. Corryn E Chini, et al. Observation of endoplasmic reticulum tubules via TOF-SIMS tandem mass spectrometry imaging of transfected cells. Biointerphases. 2018 Feb 26;13(3):03B409.  [Content Brief]

  [3]. Yun-Mi Jeong, et al. CDy6, a photostable probe for long-term real-time visualization of mitosis and proliferating cells. Chem Biol. 2015 Feb 19;22(2):299-307.  [Content Brief]

  [4]. 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.