CytoTell™ Violet 500
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Catalog Number | |
Unit Size | |
Quantity |
Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
International | See distributors |
Bulk request | Inquire |
Custom size | Inquire |
Shipping | Standard overnight for United States, inquire for international |
Molecular weight | 557.01 |
Solvent | DMSO |
Excitation (nm) | 415 |
Emission (nm) | 499 |
Intended use | Research Use Only (RUO) |
Storage | Freeze (< -15 °C); Minimize light exposure |
Overview | SDSProtocol |
Molecular weight 557.01 | Excitation (nm) 415 | Emission (nm) 499 |
Platform
Flow cytometer
Excitation | 405 nm laser |
Emission | 525/50 nm filter |
Instrument specification(s) | Pacific Orange channel |
Example protocol
AT A GLANCE
Protocol summary
- Prepare cells with test compounds
- Add 1X dye working solution
- Incubate dyes with cells at room temperature or 37 oC for 10 to 30 minutes
- Remove the dye working solution
- Analyse with flow cytometer with appropriate filter set
Important notes
Bring all the kit components at room temperature before starting the experiment. Note: The CytoTell™ dyes are lyophilized powders. They should be stable for at least 6 months if store at -20 °C, protecting from light, and avoiding freeze/thaw cycles.
Product Number |
Indicator |
Size |
Ex/Em (nm) |
Excitation Source |
22240 |
CytoTell™ UltraGreen |
500 tests |
492/519 |
488 nm (Blue Laser) |
22241 |
CytoTell™ UltraGreen |
1000 tests |
492/519 |
488 nm (Blue Laser) |
22248 |
CytoTell™ Violet 500 |
500 tests |
415/499 |
405 nm (Violet Laser) |
22251 |
CytoTell™ Blue |
500 tests |
403/454 |
405 nm (Violet Laser) |
22252 |
CytoTell™ Blue |
1000 tests |
403/454 |
405 nm (Violet Laser) |
22253 |
CytoTell™ Green |
500 tests |
511/525 |
488 nm (Blue Laser) |
22254 |
CytoTell™ Green |
1000 tests |
511/525 |
488 nm (Blue Laser) |
22255 |
CytoTell™ Red 650 |
500 tests |
628/643 |
633 nm (Red Laser) |
22256 |
CytoTell™ Red 650 |
1000 tests |
628/643 |
633 nm (Red Laser) |
22257 |
CytoTell™ Orange |
500 tests |
542 /556 |
488 nm (Blue Laser) |
22258 |
CytoTell™ Orange |
1000 tests |
542 /556 |
488 nm (Blue Laser) |
22261 |
CytoTell™ Red 590 |
500 tests |
560 /574 |
488 nm (Blue Laser) |
22262 |
CytoTell™ Red 590 |
1000 tests |
560 /574 |
488 nm (Blue Laser) |
PREPARATION OF STOCK SOLUTION
CytoTell™ dye stock solution (500X):
Add 500 µL DMSO into the dye powder vial, mix it well by vortexing to have a stock solution (500X). Note: The stock solution should be used promptly; any remaining solution should be aliquoted and frozen at < - 20 oC. Avoid repeated freeze-thaw cycles, and protect from light.
PREPARATION OF WORKING SOLUTION
CytoTellTM dye working solution (1X):
Dilute the 500X DMSO stock solution at 1 to 500 in Hanks and 20 mM Hepes buffer (HHBS) or the buffer of your choice, pH 7 (such as 1 µL of 500X DMSO stock solution to 500 µL buffer) right before use. Mix them well by vortexing. Note: The final concentration of the dye working solution should be empirically determined for different cell types and/or experimental conditions. It is recommended to test at the concentrations that are at least over ten fold range. Such as CytoTell™ Red might use much less amount in some cell types than the recommend concentrations.
SAMPLE EXPERIMENTAL PROTOCOL
- Treat cells with test compounds for a desired period of time.
- Centrifuge the cells to get 1-5 × 105 cells per tube.
- Resuspend cells in 500 µL of the CytoTell™ dye working solution. Optional: One can add the 500X DMSO stock solution into the cells directly without medium removing (such as, add 1 µL500X DMSO stock solution into 500 µL cells)
- Incubate cells with a dye solution at room temperature or 37 °C for 10 to 30 minutes, protected from light.
- Remove the dye working solution from the cells, wash the cells with HHBS or buffer of your choice. Resuspend cells in 500 µL of pre-warmed HHBS or medium to get 1-5 × 105 cells per tube.
- Monitor the fluorescence change at respected Ex/Em (see Table 1) with a flow cytometer or a fluorescence microscope.
Calculators
Common stock solution preparation
0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
1 mM | 179.53 µL | 897.65 µL | 1.795 mL | 8.976 mL | 17.953 mL |
5 mM | 35.906 µL | 179.53 µL | 359.06 µL | 1.795 mL | 3.591 mL |
10 mM | 17.953 µL | 89.765 µL | 179.53 µL | 897.65 µL | 1.795 mL |
Molarity calculator
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
/ | = | x | = |
Product Family
Name | Excitation (nm) | Emission (nm) |
CytoCalcein™ Violet 500 *Excited at 405 nm* | 420 | 505 |
ThiolTrace™ Violet 500 | 415 | 499 |
Images
Citations
Authors: Alvarez, R., Jr., Wang, B. J., Quijada, P. J., Avitabile, D., Ho, T., Shaitrit, M., Chavarria, M., Firouzi, F., Ebeid, D., Monsanto, M. M., Navarrete, N., Moshref, M., Siddiqi, S., Broughton, K. M., Bailey, B. A., Gude, N. A., Sussman, M. A.
Journal: J Mol Cell Cardiol (2019): 154-164
Authors: Jun, E. J., Song, H. Y., Park, J. H., Bae, Y. S., Paulson, B., Lee, S., Cho, Y. C., Tsauo, J., Kim, M. T., Kim, K. Y., Yang, S. G., Kim, J. K.
Journal: J Vasc Interv Radiol (2018): 1756-1763
Authors: Luan, M., Yu, L., Li, Y., Pan, W., Gao, X., Wan, X., Li, N., Tang, B.
Journal: Anal Chem (2017): 10601-10607
Authors: Zolnierowicz, J., Ambrozek-Latecka, M., Kawiak, J., Wasilewska, D., Hoser, G.
Journal: Folia Histochem Cytobiol (2013): 193-200
Authors: Begum, J., Day, W., Henderson, C., Purewal, S., Cerveira, J., Summers, H., Rees, P., Davies, D., Filby, A.
Journal: Cytometry A (2013): 1085-95
Authors: Vacaru, A. M., Isern, J., Fraser, S. T., Baron, M. H.
Journal: Genesis (2013): 751-62
Authors: He, H., Zhan, Y., Zhang, Y., Zhang, J., He, L.
Journal: Luminescence (2012): 310-4
Authors: Penjweini, R., Loew, H. G., Hamblin, M. R., Kratky, K. W.
Journal: J Microsc (2012): 100-8
Authors: Czekanska, E. M.
Journal: Methods Mol Biol (2011): 27-32
Authors: Zhang, Y., Schedle, A., Matejka, M., Rausch-Fan, X., Andrukhov, O.
Journal: Cell Mol Biol Lett (2010): 517-29
Application notes
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