CytoTell™ Red 650
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.
Spectrum
Product family
Name | Excitation (nm) | Emission (nm) |
CytoTell™ Red 590 | 562 | 587 |
Citations
Authors: Chen, Shanshan and Gong, Fusheng and Liu, Shijia and Xie, Yunqing and Ye, Xingming and Lin, Xiaowei and Wang, Xiangru and Zheng, Qiuhong and Liu, Qinying and Sun, Yang
Journal: Clinical and Experimental Medicine (2024): 204
Authors: Kubota, Sho and Sun, Yuqi and Morii, Mariko and Bai, Jie and Ideue, Takako and Hirayama, Mayumi and Sorin, Supannika and Eerdunduleng, and Yokomizo-Nakano, Takako and Osato, Motomi and others,
Journal: The EMBO Journal (2024): 1--24
Authors: Katsuyama, Naoya and Kawase, Takakazu and Barakat, Carolyne and Mizuno, M and Tomita, Akihiro and Ozeki, Kazutaka and Nishio, Nobuhiro and Sato, Yoshie and Kajiya, Ryoko and Shiraishi, Keiko and others,
Journal: Nagoya J Med Sci (2023)
Authors: Ishihara, Mikiya and Miwa, Hiroshi and Fujiwara, Hiroshi and Akahori, Yasushi and Kato, Takuma and Tanaka, Yoshimasa and Tawara, Isao and Shiku, Hiroshi
Journal: iScience (2023)
Authors: Barakat, Carolyne and Inagaki, Yuichiro and Mizuno, Shohei and Nishio, Nobuhiro and Katsuyama, Naoya and Sato, Yoshie and Kobayashi, Miki and Ozeki, Kazutaka and Iida, Hiroatsu and Tomita, Akihiro and others,
Journal: International Journal of Hematology (2023): 1--15