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Cell Meter™ IX830 fixable viability dye

Detection of Jurkat cell viability by Cell Meter™ fixable viability dye. Jurkat cells were treated and stained with Cell Meter™ IX830 (Cat#22529), and then fixed in 3.7% formaldehyde and analyzed by flow cytometry.  The dead cell population (Blue peak)  is easily distinguished from the live cell population (Red peak)  with APC-Cy7 channel, and nearly identical results were obtained before and after fixation.
Detection of Jurkat cell viability by Cell Meter™ fixable viability dye. Jurkat cells were treated and stained with Cell Meter™ IX830 (Cat#22529), and then fixed in 3.7% formaldehyde and analyzed by flow cytometry.  The dead cell population (Blue peak)  is easily distinguished from the live cell population (Red peak)  with APC-Cy7 channel, and nearly identical results were obtained before and after fixation.
Ordering information
Price ()
Catalog Number22529
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
Physical properties
SolventDMSO
Spectral properties
Excitation (nm)811
Emission (nm)822
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure
UNSPSC12171501
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OverviewpdfSDSpdfProtocol


Excitation (nm)
811
Emission (nm)
822
Discrimination and exclusion of dead cells from live cells allows cleaner separation and identification of cell populations. Cell Meter™ fixable viability dyes are a large family of cell-impermeable fluorescent viability dyes that are optimized to match the major excitation lasers of common flow cytometers, such as 350, 405, 488, 633 and 647 nm. These dyes are impermeant to live cells but permeant to cells with compromised membranes. They irreversibly react with amine- and thiol-containing proteins and other cellular components. Since dead or fixed cells with a compromised membrane more readily react with Cell Meter™ fixable cell stains, thus stain brighter than live cells with an intact membrane, these dyes can be used to assess live vs. dead status of mammalian cells. There are a few factors to be considered when using these dyes, e.g., the titration of each dye to ensure that live cells have minimal to no staining. Cell Meter™ IX830 fixable cell stain is optimized to be excited with the IR laser at 808 nm with emission at 830 nm. Compared to other commercially similar viability dyes, this fixable viability dye is much more robust and stable.

Platform


Flow cytometer

Excitation634 laser or IR lasers
Emission780/50 nm or IR filters
Instrument specification(s)APC/Cy7 or IR filters

Example protocol


AT A GLANCE

Protocol summary
  1. Prepare samples in HHBS (0.5 mL/assay)
  2. Add Cell Meter™ IX830 to the cell suspension
  3. Stain the cells at room temperature for 20 - 60 minutes
  4. Wash the cells
  5. Fix the cells (optional)
  6. Examine the sample with flow cytometer using 780/50 nm filter (APC-Cy7 channel) and/or IR filters 

Important
Thaw at room temperature before starting the experiment.

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.

Cell Meter™ IX830 stock solution (500X)
Add 200 µL of DMSO into the Cell Meter™ IX830 vial to make 500X stock solution.
Note     The unused Cell Meter™ IX830 stock solution should be divided into single use aliquots and stored at -20 °C. Avoid repeated freeze/thaw cycles.

SAMPLE EXPERIMENTAL PROTOCOL

  1. Treat cells as desired.
  2. Wash cells once with HHBS or the azide- and serum/protein-free buffer of your choice.
  3. Resuspend cells at 5 - 10 × 10 6/mL in HHBS or in the azide- and serum/protein-free buffer of your choice.
  4. Add 1 µL of 500X Cell Meter™ IX830 stock solution to 0.5 mL of cells/assay and mix it well.
  5. Incubate at room temperature, 5% CO2 incubator for 20 - 60 minutes, protected from light.
    Note     The optimal stain concentrations and incubation time should be experimentally determined for different cell lines.
  6. Wash cells twice and resuspend cells with HHBS or the buffer of your choice.
  7. Fix cells as desired (optional).
  8. Analyze cells with a flow cytometer using 780/50 nm filter (APC-Cy7 channel) or IR filters. 

Spectrum


Open in Advanced Spectrum Viewer
spectrum

Spectral properties

Excitation (nm)811
Emission (nm)822

References


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Authors: Lönnqvist, Susanna and Junker, Johan P E and Sedell, Maria and Nyman, Erika and Kratz, Gunnar
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Influence of traditionally used Nepalese plants on wound healing and immunological properties using primary human cells in vitro.
Authors: Zimmermann-Klemd, Amy M and Konradi, Viktoria and Steinborn, Carmen and Ücker, Annekathrin and Falanga, Chiara Madlen and Woelfle, Ute and Huber, Roman and Jürgenliemk, Guido and Rajbhandari, Meena and Gründemann, Carsten
Journal: Journal of ethnopharmacology (2019): 415-423
Evaluation of the Compatibility of Electric Tumor Treating Fields with Key Anti-tumoral T-Cell Functions.
Authors: Simchony, Hadar and Diamant, Diamant and Ram, Zvi and Volovitz, Ilan
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