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FerroBrite™ Green
Example protocol
AT A GLANCE
Before initial use, thaw FerroBrite™ Green at room temperature and briefly centrifuge to collect the dried pellet.
Treat the cells as desired.
Remove the treatment and add 100 µL of FerroBrite™ Green working solution.
Incubate at 37 °C for 30 minutes.
Use an Ex/Em = 450/550 filter set to monitor the fluorescence signal.
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
Prepare a 5 to 10 mM FerroBrite™ Green stock solution in DMSO. For example, to make a 10 mM FerroBrite™ Green stock solution add 284 µL of DMSO to the vial of FerroBrite™ Green, and mix thoroughly.
Note: Prepare single-use aliquots of any remaining FerroBrite™ Green stock solution. Store aliquots at ≤ -20 º C, protected from light. Avoid freeze/thaw cycles.
PREPARATION OF WORKING SOLUTION
Prepare a 10 to 20 μM working solution by diluting the FerroBrite™ Green stock solution into Hanks' solution with 20 mM Hepes buffer (HHBS, AAT Cat No. 20011).
Note: For optimal results, use this solution within 2 hours of preparation.
Note: Protect the working solution from light by covering it with foil or placing it in the dark.
SAMPLE EXPERIMENTAL PROTOCOL
Plate the cells as desired in a 96-well black wall-clear bottom plate.
Add the drug of interest to the cells at the necessary concentration.
Note: For a positive control, add Erastin at a concentration of 10 µM to the cells. Incubate the cells with Erastin for 5 to 6 hours at 37°C in a 5% CO2 incubator to induce ferroptosis.
Remove the cell culture medium and add 100 µL of FerroBrite™ Green working solution to the cells.
Incubate the cells at 37°C for 20 to 30 minutes, keeping them protected from light.
Note: The optimal concentration and incubation time for FerroBrite™ Green may vary depending on the cell line used. It is recommended to test various concentrations to determine the most effective dose.
Remove the FerroBrite™ Green working solution and wash the cells twice using HHBS buffer.
Add HHBS buffer and analyze the cells using a fluorescence microscope with an Ex/Em = 450/550 nm filter set.
Spectrum
Product family
| Name | Excitation (nm) | Emission (nm) |
| Indocyanine Green *CAS 3599-32-4* | 789 | 813 |
| Helixyte™ Green *20X Aqueous PCR Solution* | 498 | 522 |
| Helixyte™ Green *10,000X Aqueous PCR Solution* | 498 | 522 |
| Thiolite™ Green | 505 | 524 |
| CytoTell™ Green | 510 | 525 |
| LysoBrite™ Green | 501 | 510 |
| Q4ever™ Green *1250X DMSO Solution* | 503 | 527 |
| Droplite™ Green | 421 | 521 |
References
Authors: Zhao, Zhao and Jin, Wendong and Wu, Mengfan and Lin, Qingyu and Duan, Yixiang
Journal: Chemical communications (Cambridge, England) (2024): 7773-7776
Authors: Zhong, Lili and Fu, Datian and Xu, Jin and Tan, Linyan and Wu, Haimei and Wang, Min
Journal: RSC advances (2024): 12864-12872
Authors: Che, Yiran and Yang, Jingying and Dong, Zhenming and Wang, Jianhua and Yan, Xiaoqing and Wang, Yu and Shuang, Shaomin
Journal: Spectrochimica acta. Part A, Molecular and biomolecular spectroscopy (2024): 123799
Authors: Mehmood, Abdul Hadi and Chang, Jia and Wang, Yan and Li, Shijing and Ma, Jiale and Dong, Baoli and Liu, Hong
Journal: Analytical methods : advancing methods and applications (2024): 3486-3491
Authors: Liu, Ruixin and Jiang, Haijing and Yang, Wenjie and Zheng, Zhijuan and Wang, Xiaoming and Tian, Zhenhua and Wang, Danyang and Kan, Dongfang and Zhang, Dan and Tang, Zhixin
Journal: Analytica chimica acta (2024): 342673
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