Cell Navigator® Live Cell Endoplasmic Reticulum (ER) Staining Kit Red Fluorescence

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Telephone	1-800-990-8053
Fax	1-800-609-2943
Email	sales@aatbio.com
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Shipping	Standard overnight for United States, inquire for international

Spectral properties

Excitation (nm)	588
Emission (nm)	620

Storage, safety and handling

Certificate of Origin	Download PDF
H-phrase	H303, H313, H333
Hazard symbol	XN
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R22
UNSPSC	12352200

Alternative formats

Cell Navigator® Live Cell Endoplasmic Reticulum (ER) Staining Kit *Blue Fluorescence*

Cell Navigator® Live Cell Endoplasmic Reticulum (ER) Staining Kit *Green Fluorescence*

Overview SDS Protocol

Excitation (nm)

588

Emission (nm)

620

The endoplasmic reticulum (ER) is a type of organelle in the cells of eukaryotic organisms that forms an interconnected network of flattened, membrane-enclosed sacs or tube-like structures known as cisternae. The membranes of the ER are continuous with the outer nuclear membrane. ER occurs in most types of eukaryotic cells, but is absent from red blood cells and spermatozoa. This Cell Navigator® Live Cell Endoplasmic Reticulum (ER) Staining Kit uses our ER Tracer™ Red as an ER marker. ER Tracer™ Red stain is a cell-permeant fluorescent dye that is highly selective for ER. This stain consists of a red fluorescent dye and ER binder that selectively bind to ER in most of cell types. For some cells, ER Tracer™ Red may not selectively bind to ER. ER Tracer™ Red has spectral properties essentially identical to Texas Red, making this kit convenient with the Texas Red filter set.

Platform

Fluorescence microscope

Excitation	590 nm
Emission	620 nm
Recommended plate	Black wall/clear bottom
Instrument specification(s)	TRITC or Cy3 filter

Components

Example protocol

AT A GLANCE

Protocol Summary

Prepare cells in growth medium
Incubate cells with ER Tracer™ Red working solution at 37 °C for 15 - 30 minutes
Analyze under fluorescence microscope at Ex/Em = 590/620 nm (TRITC or Cy3 filter set)

Important Thaw all the kit components at room temperature before starting the experiment.

CELL PREPARATION

For guidelines on cell sample preparation, please visit https://www.aatbio.com/resources/guides/cell-sample-preparation.html

PREPARATION OF STOCK SOLUTIONS

Unless otherwise noted, all unused stock solutions should be divided into single-use aliquots and stored at -20 °C after preparation. Avoid repeated freeze-thaw cycles.

ER Tracer™ Red stock solution (500X)

Add 20 µL of DMSO (Component C) into the vial of ER Tracer™ Red (Component A) and mix well to make 500X ER Tracer™ Red stock solution.
Note 20 µL of 500X ER Tracer™ Red stock solution is enough for one 96-well plate. Unused 500X ER Tracer™ Red stock solution can be stored at ≤ -20 ºC for two weeks if the tubes are sealed tightly. Protect from light.

PREPARATION OF WORKING SOLUTION

ER Tracer™ Red working solution

Add 20 µL of 500X ER Tracer™ Red stock solution into 10 mL of Live Cell Staining Buffer (Component B), and mix well to make ER Tracer™ Red working solution.
Note This ER Tracer™ Red working solution is stable for at least 2 hours at room temperature. Protect from light.

SAMPLE EXPERIMENTAL PROTOCOL

Add 100 µL/well (96-well plate) or 50 µL/well (384-well plate) of ER Tracer™ Red working solution in the cell plate. Incubate cells with working solution at 37 °C for 15 - 30 minutes, protected from light.
Note The optimal concentration of the ER probe varies depending on the specific application. Concentration higher than the working solution can be toxic to cells. The staining conditions may be modified according to the particular cell type and the permeability of the cells or tissues to the probe.
Remove ER Tracer™ Red working solution in each well. Wash cells with physically relevant buffer three times.
Fix cells after staining (Optional). Fix the cells with 4% formaldehyde for 5 - 10 minutes. Wash cells with physically relevant buffer three times.
Observe the fluorescence signal in cells using fluorescence microscope with TRITC or Cy3 filter set (Ex/Em = 590/620 nm).

Spectrum

Open in Advanced Spectrum Viewer

Spectral properties

Excitation (nm)	588
Emission (nm)	620

Product Family

Name	Excitation (nm)	Emission (nm)
Cell Navigator® Live Cell Endoplasmic Reticulum (ER) Staining Kit Blue Fluorescence	344	457
Cell Navigator® Live Cell Endoplasmic Reticulum (ER) Staining Kit Green Fluorescence	503	511

Images

Figure 1. Fluorescence images of endoplasmic reticulum (ER) staining in HeLa cells cultured in a 96-well black-wall clear-bottom plate using a fluorescence microscope equipped with a TRITC filter set. Live cells were stained with Cell Navigator® Live Cell Endoplasmic Reticulum (ER) Staining Kit *Red Fluorescence* (Cat#22636, Red), mitochondria dye MitoLite™ Green (Cat#22675, Green), and nuclei stain Hoechst 33342 (Cat#17530, Blue).

Figure 2. Fluorescence images of endoplasmic reticulum (ER) staining in HeLa cells cultured in a 96-well black-wall clear-bottom plate using a fluorescence microscope equipped with a TRITC filter set. Live cells were stained with Cell Navigator® Live Cell Endoplasmic Reticulum (ER) Staining Kit *Red Fluorescence* (Cat#22636, Red) then fixed with 4% formaldehyde and labeled with F-actin dye iFluor® 488-Phalloidin (Cat#22661, Green) and nuclei stain DAPI (Cat#17507, Blue).