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

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).
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).
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).
Ordering information
Price ()
Catalog Number22636
Unit Size
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Additional ordering information
Telephone1-408-733-1055
Fax1-408-733-1304
Emailsales@aatbio.com
InternationalSee distributors
ShippingStandard overnight for United States, inquire for international
Spectral properties
Excitation (nm)588
Emission (nm)620
Storage, safety and handling
Certificate of OriginDownload PDF
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
UNSPSC12352200
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OverviewpdfSDSpdfProtocol


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

Excitation590 nm
Emission620 nm
Recommended plateBlack wall/clear bottom
Instrument specification(s)TRITC or Cy3 filter

Components


Component A: ER Tracer™ Red1 vial
Component B: Live Cell Staining Buffer1 bottle (20 mL)
Component C: DMSO1 vial (100 µL)

Example protocol


AT A GLANCE

Protocol Summary
  1. Prepare cells in growth medium
  2. Incubate cells with ER Tracer™ Red working solution at 37 °C for 15 - 30 minutes
  3. 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

  1. 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.
  2. Remove ER Tracer™ Red working solution in each well. Wash cells with physically relevant buffer three times.
  3. Fix cells after staining (Optional). Fix the cells with 4% formaldehyde for 5 - 10 minutes. Wash cells with physically relevant buffer three times.
  4. Observe the fluorescence signal in cells using fluorescence microscope with TRITC or Cy3 filter set (Ex/Em = 590/620 nm). 

Spectrum


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spectrum

Spectral properties

Excitation (nm)588
Emission (nm)620