InCellGen™ ER Tracker Green£¬IC-1560

Description

ER-Tracker dyes are cell-permeant, live-cell stains that are highly selective for the endoplasmic reticulum (ER). These dyes rarely stain mitochondria, unlike the conventional ER stain DiOC6(3), and staining at low concentrations does not appear to be toxic to cells. When cells are stained using the optimized protocol provided, staining patterns are partially retained after

treatment with formaldehyde.

Experimental Protocol

This protocol was optimized using bovine pulmonary artery endothelial cells and has been confirmed in other common cell lines. Recommendations for experimental protocols should be

used as a starting point, and optimal labeling conditions for each cell type should be determined empirically.

Reagent Preparation

ER-Tracker Blue-White DPX is supplied as aliquots of a 1 mM stock solution in DMSO. Allow each vial to warm to room temperature before use, then briefly centrifuge to deposit the DMSO solution at the bottom of the vial.

ER-Tracker Green and ER-Tracker Red dyes are supplied as 100ug oflyophilized material. Prepare a 1 mM stock solution of the appropriate dye: for ER-Tracker Green dye, dissolve the

contents of the vial in 128 ¦ÌL of DMSO; for ER-Tracker Red dye, dissolve the contents of the vial in 110ul of DMSO. It is recommended that the 1 mM solution then be separated into aliquots and stored frozen with desiccant.

Cell Preparation and Staining

1.Prepare staining solution. Dilute the 1 mM stock solution to the final working concentration. We recommend working  concentrations of 1uM for ER-Tracker™ Green and ER-Tracker™Red dyes. To minimize potential labeling artifacts, use the low-est dye concentrations possible. Best results are obtained when staining is performed in Hank,s Balanced Salt Solution with calcium and magnesium (HBSS/Ca/Mg,) at37¡æ/ 5% CO2. 

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¨C30 minutes at 37¡æ. 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.

Storage  

Store at ¨C20¡æ£¬Avoid freeze-thaw cycles£¬Protect from light.

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