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Cell Meter™ VX500 fixable viability dye

Detection of Jurkat cell viability by Cell Meter™ fixable viability dye. Jurkat cells were treated and stained with Cell Meter™ VX500 (Cat#22542), 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 Pacific Orange 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™ VX500 (Cat#22542), 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 Pacific Orange channel, and nearly identical results were obtained before and after fixation.
Ordering information
Price ()
Catalog Number22542
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)433
Emission (nm)498
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)
433
Emission (nm)
498
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™ VX500 fixable cell stain is optimized to be excited with the violet laser at 405 nm with emission at 500 nm. Compared to other commercially similar viability dyes, this fixable viability dye is much more robust and stable.

Platform


Flow cytometer

Excitation405 nm laser
Emission525/50 nm filter
Instrument specification(s)Pacific Orange channel

Example protocol


AT A GLANCE

Protocol summary
  1. Prepare samples in HHBS (0.5 mL/assay)
  2. Add Cell Meter™ VX500 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 525/50 nm filter (Pacific Orange channel) 

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™ VX500 stock solution (500X)
Add 200 µL of DMSO into the Cell Meter™ VX500 vial to make 500X stock solution.
Note     The unused Cell Meter™ VX500 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™ VX500 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 525/50 nm filter (Pacific Orange channel). 

Spectrum


Open in Advanced Spectrum Viewer
spectrum

Spectral properties

Excitation (nm)433
Emission (nm)498

References


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PGC-1α Regulates Cell Proliferation and Invasion via AKT/GSK-3β/β-catenin Pathway in Human Colorectal Cancer SW620 and SW480 Cells.
Authors: Yun, Seong-Hoon and Park, Joo-In
Journal: Anticancer research (2020): 653-664
CFSE dilution to study human T and NK cell proliferation in vitro.
Authors: Terrén, Iñigo and Orrantia, Ane and Vitallé, Joana and Zenarruzabeitia, Olatz and Borrego, Francisco
Journal: Methods in enzymology (2020): 239-255
The miR-3940-5p inhibits cell proliferation of gingival mesenchymal stem cells.
Authors: Han, Xiao and Yang, Haoqing and Cao, Yangyang and Ge, Lihua and Han, Nannan and Zhang, Chen and Fan, Zhipeng and Yao, Rui
Journal: Oral diseases (2019): 1363-1373
Toll-Like Receptor 4 (TLR4)/Cyclooxygenase-2 (COX-2) Regulates Prostate Cancer Cell Proliferation, Migration, and Invasion by NF-κB Activation.
Authors: Wang, Wei and Wang, Jiye
Journal: Medical science monitor : international medical journal of experimental and clinical research (2018): 5588-5597
Evaluation of Cell Proliferation and Apoptosis in Immunotoxicity Testing.
Authors: Nagarkatti, Mitzi and Rieder, Sadiye Amcaoglu and Nagarkatti, Prakash S
Journal: Methods in molecular biology (Clifton, N.J.) (2018): 209-230
Quantitative analysis of cell proliferation by a dye dilution assay: Application to cell lines and cocultures.
Authors: Chung, Soobin and Kim, Seol-Hee and Seo, Yuri and Kim, Sook-Kyung and Lee, Ji Youn
Journal: Cytometry. Part A : the journal of the International Society for Analytical Cytology (2017): 704-712
[Myeloid-derived suppressor cells promoted autologous B cell proliferation in rheumatoid arthritis].
Authors: Chen, W and Hu, F L and Liu, H J and Xu, L L and Li, Y N and Li, Z G
Journal: Beijing da xue xue bao. Yi xue ban = Journal of Peking University. Health sciences (2017): 819-823
Gag-Specific CD4 T Cell Proliferation, Plasmacytoid Dendritic Cells, and Ethnicity in Perinatally HIV-1-Infected Youths: The ANRS-EP38-IMMIP Study.
Authors: Scott-Algara, Daniel and Warszawski, Josiane and Chenadec, Jérôme Le and Didier, Céline and Montange, Thomas and Viard, Jean-Paul and Dollfus, Catherine and Avettand-Fenoel, Véronique and Rouzioux, Christine and Blanche, Stéphane and Buseyne, Florence
Journal: AIDS research and human retroviruses (2017): 21-28
Hypoxia inhibits mesenchymal stem cell proliferation through HIF1α-dependent regulation of P27.
Authors: Kumar, Sanjay and Vaidya, Meenal
Journal: Molecular and cellular biochemistry (2016): 29-38
Vanadium pentoxide prevents NK-92MI cell proliferation and IFNγ secretion through sustained JAK3 phosphorylation.
Authors: Gallardo-Vera, Francisco and Diaz, Daniel and Tapia-Rodriguez, Miguel and Fortoul van der Goes, Teresa and Masso, Felipe and Rendon-Huerta, Erika and Montaño, Luis F
Journal: Journal of immunotoxicology (2016): 27-37