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Cell Navigator® Live Cell Endoplasmic Reticulum (ER) Staining Kit *Blue 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 with a DAPI filter set. (A) Live cells were co-stained with ER-selective probe ER Tracer™ Blue (Cat No. 22634, Blue) and Nuclear Red™ LCS1 (Cat No. 17542, Red). (B) Live cells were co-stained with ER-selective probe ER Tracer™ Blue (Cat No. 22634, Blue) and Nuclear Red™ LCS1 (Cat No. 17542, Red) and then were fixed with 4% formaldehyde.
Fluorescence images of endoplasmic reticulum (ER) staining in HeLa cells cultured in a 96-well black-wall clear-bottom plate using a fluorescence microscope with a DAPI filter set. (A) Live cells were co-stained with ER-selective probe ER Tracer™ Blue (Cat No. 22634, Blue) and Nuclear Red™ LCS1 (Cat No. 17542, Red). (B) Live cells were co-stained with ER-selective probe ER Tracer™ Blue (Cat No. 22634, Blue) and Nuclear Red™ LCS1 (Cat No. 17542, Red) and then were fixed with 4% formaldehyde.
Ordering information
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Catalog Number22634
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)344
Emission (nm)457
Storage, safety and handling
Certificate of OriginDownload PDF
Intended useResearch Use Only (RUO)
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OverviewpdfSDSpdfProtocol


Excitation (nm)
344
Emission (nm)
457
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™ Blue as an ER marker. ER Tracer™ Blue stain is a cell-permeant fluorescent dye that is highly selective for ER. This stain consists of a blue fluorescent dye and ER binder that selectively bind to ER in most of cell types. For some cells, ER Tracer™ Blue may not selectively bind to ER. ER Tracer™ Blue has spectral properties similar to DAPI, making this kit convenient with the DAPI filter set.

Platform


Fluorescence microscope

ExcitationDAPI filter set
EmissionDAPI filter set
Recommended plateBlack wall/clear bottom

Components


Component A: ER Tracer™ Blue1 vial
Component B: Live Cell Staining Buffer1 bottle (20 mL)
Component C: DMSO1 vial (100 uL)

Example protocol


AT A GLANCE

Protocol Summary
  1. Prepare cells in growth medium
  2. Incubate cells with ER Tracer™ Blue working solution at 37 oC for 15 - 30 minutes
  3. Analyze under fluorescence microscope with DAPI 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™ Blue stock solution (500X)
Add 20 µL of DMSO (Component C) into ER Tracer™ Blue (Component A) to make 500X stock solution.

PREPARATION OF WORKING SOLUTION

ER Tracer™ Blue working solution
Add 2 µL of 500X stock solution into 1 mL of Live Cell Staining Buffer (Component B), and mix well. The working solution is stable for at least 2 hours at room temperature. 
Note     20 µL of 500X ER Tracer™ Blue stock solutions is enough for one 96-well plate. Unused ER Tracer™ Blue 500X stock solution can be aliquoted and stored at ≤ -20 oC for two weeks if the tubes are sealed tightly. Protect from light and avoid repeated freeze-thaw cycles.

SAMPLE EXPERIMENTAL PROTOCOL

  1. Treat samples as desired.
    Note     Working solution can be added directly into the cell culture medium. Alternatively, remove the cell culture medium and wash with the buffer of your choice.
  2. Add 100 µL/well (96-well plate) or 50 µL/well (384-well plate) of ER Tracer™ Blue working solution in the cell plate. Incubate cells with working solution at 37 oC 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.
  3. Remove working solution in each well. Wash cells with physically relevant buffer three times.
  4. Fix cells after staining (Optional). Fix the cells with 4% formaldehyde for 5 -10 minutes. Wash cells with physically relevant buffer three times.
  5. Observe the fluorescence signal in cells using fluorescence microscope with a DAPI filter set. 

Spectrum


Open in Advanced Spectrum Viewer
spectrum

Spectral properties

Excitation (nm)344
Emission (nm)457