Cell Meter™ Fluorimetric Intracellular Total ROS Activity Assay Kit*Green Fluorescence*
|Shipping||Standard overnight for United States, inquire for international|
|H-phrase||H303, H313, H333|
|Intended use||Research Use Only (RUO)|
|R-phrase||R20, R21, R22|
|Recommended plate||Black wall/clear bottom|
Fluorescence microplate reader
|Recommended plate||Black wall/clear bottom|
|Instrument specification(s)||Bottom read mode|
AT A GLANCE
- Prepare cells in growth medium
- Add Amplite™ ROS Green working solution (100 µL/well for a 96- well plate or 25 µL/well for a 384-well plate)
- Stain the cells at 37°C for 60 minutes
- Treat the cells with test compounds to induce ROS
- Monitor the fluorescence increase (bottom read mode) at Ex/Em= 490/525 nm (Cutoff = 515 nm) or fluorescence microscope with FITC filter set
Thaw all the kit components at room temperature before starting the experiment.
PREPARATION OF STOCK SOLUTION
1. Amplite™ ROS Green stock solution (500X):
Add 40 µL of DMSO (Component C) into the vial of Amplite™ ROS Green (Component A) and mix well to make 500X Amplite™ ROS Green stock solution. Protect from light. Note: 20 µL of 500X Amplite™ ROS Green stock solution is enough for 1 plate. For storage, seal tubes tightly.
PREPARATION OF WORKING SOLUTION
Add 20 µL of 500X Amplite™ ROS Green stock solution into 10 mL of Assay Buffer (Component B) and mix well to make Amplite™ ROS Green working solution. Note: This Amplite™ ROS Green working solution is stable for at least 2 hours at room temperature.
For guidelines on cell sample preparation, please visit
SAMPLE EXPERIMENTAL PROTOCOL
- Add 100 µL/well (96-well plate) or 25 µL/well (384-well plate) of Amplite™ ROS Green working solution into the cell plate.
- Incubate the cells in a 5% CO2, 37°C incubator for one hour.
- Treat cells with 20 µL of 11X test compounds (96-well plate) or 10 µL of 6X test compounds (384-well plate) in your desired buffer (such as PBS or HHBS). For control wells (untreated cells), add the corresponding amount of compound buffer.
- To induce ROS, incubate the cell plate at room temperature or in a 5% CO2, 37°C incubator for at least 15 minutes or a desired period of time (30 minutes for Hela cells treated with 1 mM H2O2).
- Monitor the fluorescence increase with a fluorescence microplate reader (bottom read mode) at Ex/Em = 490/525 nm (Cutoff = 515 nm) or observe cells using a fluorescence microscope with FITC filter set.
Authors: Li, Xiaoming and Zhang, Xiang and Hou, Xiaozhi and Bing, Xin and Zhu, Fangyuan and Wu, Xinhao and Guo, Na and Zhao, Hui and Xu, Fenglei and Xia, Ming
Journal: Apoptosis (2022): 1--15
Authors: Tucker, Katherine and Yin, Yajie and Staley, Stuart-Allison and Zhao, Ziyi and Fang, Ziwei and Fan, Yali and Zhang, Xin and Suo, Hongyan and Sun, Wenchuan and Prabhu, Varun Vijay and others,
Journal: American Journal of Cancer Research (2022): 521
Authors: Teng, Yu-Ning and Hung, Chin-Chuan and Kao, Pei-Heng and Chang, Ying-Tzu and Lan, Yu-Hsuan
Journal: Biomedicine \& Pharmacotherapy (2022): 113832
Authors: Li, Jing and Qi, Fangzheng and Su, Huishan and Zhang, Chuanshan and Zhang, Qing and Chen, Ying and Chen, Ping and Su, Linjia and Chen, Yanan and Yang, Yuqi and others,
Journal: International journal of biological sciences (2022): 2914
Authors: Cai, Dong-Mei and Mei, Fan-Biao and Zhang, Chao-Jun and An, San-Chun and Lv, Rui-Bo and Ren, Guan-Hua and Xiao, Chan-Chan and Long, Long and Huang, Tian-Ren and Deng, Wei
Journal: Journal of Hepatocellular Carcinoma (2022): 1229--1246
Authors: Jiang, Xuan and Zhang, Xin and Fu, Chao and Zhao, Ruili and Jin, Tianming and Liu, Mengyue and Pan, Chenhao and Li, Liu An and Ma, Jifei and Yu, Enyuan and others,
Journal: The Protein Journal (2021): 1--12
Authors: Tang, Jingshu and Kang, Yuying and Zhou, Yujun and Li, Xinnan and Lan, Jiaqi and Wu, Lei and Feng, Xinhong and Peng, Ying
Journal: Neurobiology of Disease (2021): 105315
Authors: Shimoda, Ayumu and Tanabe, Takemi and Sato, Tsubasa and Nedachi, Taku
Journal: Bioscience, Biotechnology, and Biochemistry (2021): 2103--2112
Authors: Teng, Yu-Ning and Wang, Charles CN and Liao, Wei-Chieh and Lan, Yu-Hsuan and Hung, Chin-Chuan
Journal: Molecules (2020): 247
Authors: Lakshmi, Sowmya P and Reddy, Aravind T and Kodidhela, Lakshmi Devi and Varadacharyulu, N Ch
Journal: Life Sciences (2020): 118260
Authors: Magalhaes LM, Lucio M, Segundo MA, Reis S, Lima JL.
Journal: Talanta (2009): 1219
Authors: Amaral S, Oliveira PJ, Ramalho-Santos J.
Journal: Curr Diabetes Rev (2008): 46
Authors: Murray BK, Ohmine S, Tomer DP, Jensen KJ, Johnson FB, Kirsi JJ, Robison RA, O'Neill KL.
Journal: J Virol Methods (2008): 74
Authors: Starkov AA., undefined
Journal: Ann N Y Acad Sci (2008): 37
Authors: Wada M., undefined
Journal: Yakugaku Zasshi (2008): 1031
Authors: Perrone GG, Tan SX, Dawes IW.
Journal: Biochim Biophys Acta (2008): 1354
Authors: Armstrong JS, Whiteman M.
Journal: Methods Cell Biol (2007): 355
Authors: Zhang W, Wang M, Xie HY, Zhou L, Meng XQ, Shi J, Zheng S.
Journal: Transplant Proc (2007): 1332
Authors: Gorlach A, Kietzmann T.
Journal: Methods Enzymol (2007): 421
Authors: Afonso V, Champy R, Mitrovic D, Collin P, Lomri A.
Journal: Joint Bone Spine (2007): 324
A Comparison of Fluorescent Red Calcium Indicators for Detecting Intracellular Calcium Mobilization in CHO Cells
A Meta-Analysis of Common Calcium Indicators
A New Red Fluorescent & Robust Screen Quest™ Rhod-4™ Ca2+Indicator for Screening GPCR & Ca2+ Channel Targets
A New Robust No-Wash FLIPR Calcium Assay Kit for Screening GPCR and Calcium Channel Targets