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Cell Meter™ VX500 fixable viability dye
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.
Example protocol
AT A GLANCE
Protocol summary
- Prepare samples in HHBS (0.5 mL/assay)
- Add Cell Meter™ VX500 to the cell suspension
- Stain the cells at room temperature for 20 - 60 minutes
- Wash the cells
- Fix the cells (optional)
- 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.
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.
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
- Treat cells as desired.
- Wash cells once with HHBS or the azide- and serum/protein-free buffer of your choice.
- Resuspend cells at 5 - 10 × 10 6/mL in HHBS or in the azide- and serum/protein-free buffer of your choice.
- Add 1 µL of 500X Cell Meter™ VX500 stock solution to 0.5 mL of cells/assay and mix it well.
- 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. - Wash cells twice and resuspend cells with HHBS or the buffer of your choice.
- Fix cells as desired (optional).
- Analyze cells with a flow cytometer using 525/50 nm filter (Pacific Orange channel).
Spectrum
Product family
| Name | Excitation (nm) | Emission (nm) |
| Cell Meter™ BX520 fixable viability dye | 491 | 516 |
| Cell Meter™ BX590 fixable viability dye | 492 | 579 |
| Cell Meter™ BX650 fixable viability dye | 518 | 654 |
| Cell Meter™ IX830 fixable viability dye | 811 | 822 |
| Cell Meter™ RX660 fixable viability dye | 649 | 664 |
| Cell Meter™ RX700 fixable viability dye | 690 | 713 |
| Cell Meter™ RX780 fixable viability dye | 629 | 767 |
| Cell Meter™ VX450 fixable viability dye | 406 | 445 |
References
View all 50 references: Citation Explorer
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
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
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
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
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
Authors: Nagarkatti, Mitzi and Rieder, Sadiye Amcaoglu and Nagarkatti, Prakash S
Journal: Methods in molecular biology (Clifton, N.J.) (2018): 209-230
Page updated on November 21, 2024
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