ER-Tracker Green

Reagent Preparation

ER-Tracke Green and ER-Tracker Red dyes are supplied as lyophilized material. Prepare a 1 mM stock solution of the appropriate dye: for ER-Tracker Green dye, dissolve 100 μg the contents of the vial in 128 μL of DMSO. It is recommended that the 1 mM solution then be separated into aliquots and stored frozen with desiccant，Keep away from light.

Cell Preparation and Staining

1.1 Prepare staining solution. Dilute the 1 mM stock solution to the final working concentration. We recommend working concentrations of 1 μM for ER-Tracker Green. To minimize potential labeling artifacts, use the lowest dye concentrations possible. Best results are obtained when staining is performed in Hankʼs Balanced Salt Solution with calcium and magnesium at 37˚C/5% CO₂.

1.2 Stain the cells. For adherent cells, remove the medium from the culture dish, rinse with HBSS, and add prewarmed staining solution. Incubate the cells for approximately 15–30 minutes at 37˚C. Replace the staining solution with fresh probe-free medium and view the cells using a fluorescence microscope. If the stained cells are to be fixed, refer to the fixation steps below for the appropriate dye.

Fixation for ER-Tracker Green Dye

3.1 Fix cells. If stained cells are to be fixed, fixation is recommended using 4% formaldehyde for 2 minutes at 37˚C.

3.2 Wash and view cells. After fixation, perform two 5-minute washes in any suitable buffer prior to mounting, viewing, or further staining. Permeabilization is not recommended; signal is not retained after permeabilization with Triton X-100.

Determination for ER-Tracker Green Dye

ER-Tracker Green showed green fluorescence, with a maximum excitation wavelength of 504nm and a maximum emission wavelength of 511nm.