ER-Tracker Green 

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 cells after fixation^[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 128 μL DMSO to obtain a 1 mM stock solution.
Note: It is recommended to store the stock solution at -20°C or -80°C, protected from light, and avoid repeated freeze-thaw cycles.
1.2 Preparation of ER-Tracker working solution
Dilute the stock solution with serum-free cell culture medium or PBS to obtain a 100 nM-1 μM working solution.
Note: The concentration of the ER-Tracker working solution should be adjusted according to actual conditions.
2. Cell staining
2.1 Suspension cells (6-well plate)
a. Centrifuge at 1000 g at 4°C for 3-5 minutes, discard the supernatant. Wash twice with PBS, 5 minutes each time. Adjust the cell density to 1×10⁶/mL.
b. Add 1 mL working solution and incubate at room temperature for 5-30 minutes.
c. Centrifuge at 400 g at 4°C for 3-4 minutes and discard the supernatant.
d. Wash twice with PBS, 5 minutes each time.
e. Resuspend the cells in serum-free cell culture medium or PBS. Observe using a fluorescence microscope or flow cytometry.
2.2 Adherent cells
a. Culture adherent cells on sterile coverslips.
b. Remove the coverslips from the culture medium and aspirate excess medium.
c. Add 100 μL working solution, gently shake to fully cover the cells, and incubate at room temperature for 5-30 minutes.
d. Wash twice with culture medium, 5 minutes each time. Observe using a fluorescence microscope or flow cytometer.
Note: If flow cytometry is used, cells need to be resuspended before staining.

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

783.09

C₃₇H₄₂BClF₂N₆O₆S

520

489

730931-46-1

Solid

Brown to reddish brown

O=C(NS(=O)(C1=CC=C(CCNC(C2=C(OC)C(NC(CCC3=CC=C4C=C5C(C)=CC(C)=[N]5[B+3]([F-])([N-]43)[F-])=O)=CC(Cl)=C2)=O)C=C1)=O)NC6CCCCC6

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]. 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.

  [3]. J Jacob Strouse, et al. Fluorescent substrates for flow cytometric evaluation of efflux inhibition in ABCB1, ABCC1, and ABCG2 transporters. Anal Biochem. 2013 Jun 1;437(1):77-87.  [Content Brief]