Cell Navigator® Mitochondrion Staining Kit *Green Fluorescence*
Overview | ![]() ![]() |
Excitation (nm) 508 | Emission (nm) 528 |
Platform
Fluorescence microscope
Excitation | FITC filter set |
Emission | FITC filter set |
Recommended plate | Black wall/clear bottom |
Components
Component A: MitoLite™ Green | 1 vial (100 µL, 500X DMSO stock solution) |
Component B: Live Cell Staining Buffer | 1 bottle (50 mL) |
Example protocol
AT A GLANCE
Protocol summary
- Prepare cells
- Add Mitolite™ Green working solution
- Incubate at 37°C for 30 minutes to 2 hours
- Analyze the cells under fluorescence microscope at Ex/Em = 485/525 nm (FITC filter set)
Important notes
Thaw all the components at room temperature before starting the experiment.
PREPARATION OF WORKING SOLUTION
Add 20 µL of 500X Mitolite™ Green (Component A) into 10 mL of Live Cell Staining Buffer (Component B) to make Mitolite™ Green working solution. Protect from light. Note: 20 µL of 500X Mitolite™ Green (Component A) is enough for one 96-well plate. The optimal concentration of the fluorescent mitochondrial indicator varies depending on the specific application. The staining conditions may be modified according to the particular cell type and the permeability of the cells or tissues to the probe.
For guidelines on cell sample preparation, please visit
https://www.aatbio.com/resources/guides/cell-sample-preparation.html
SAMPLE EXPERIMENTAL PROTOCOL
For adherent cells:
- Grow cells either in a 96-well black wall/clear bottom plate (100 µL/well/96-well plate) or on cover-slips inside a petri dish filled with the appropriate culture medium.
- When cells reach the desired confluence, add equal volume of Mitolite™ Green working solution.
- Incubate the cells in a 37°C, 5% CO2 incubator for 30 minutes to 2 hours.
- Replace Mitolite™ Green working solution with Hanks and 20 mM Hepes buffer (HHBS) or buffer of your choice (e.g. the buffer with growth medium at 1:1 concentration).
- Observe the cells using a fluorescence microscope with FITC filter set (Ex/Em = 485/525 nm). Note: It’s recommended to increase either the labeling concentration or the incubation time to allow the dye to accumulate if the cells do not appear to be sufficiently stained.
For suspension cells:
- Centrifuge the cells at 1000 rpm for 5 minutes to obtain a cell pellet and aspirate the supernatant.
- Resuspend the cell pellets gently in pre-warmed (37°C) growth medium, and add equal volume of Mitolite™ Green working solution.
- Incubate the cells in a 37°C, 5% CO2 incubator for 30 minutes to 2 hours.
- Replace Mitolite™ Green working solution with Hanks and 20 mM Hepes buffer (HHBS) or buffer of your choice (e.g. the buffer with growth medium at 1:1 concentration).
- Observe the cells using a fluorescence microscope with FITC filter set (Ex/Em = 485/525 nm). Note: It is recommended to increase either the labeling concentration or the incubation time to allow the dye to accumulate if the cells do not appear to be sufficiently stained. Suspension cells may be attached to cover-slips that have been treated with BD Cell-Tak® (BD Biosciences) and stained as adherent cells.
Product family
Images
Citations
Authors: Zhang, Jiao Jiao and Wang, Xian Zhong and Do, Huynh Luong and Ch, undefined and imali, Nisansala and Kang, Tae Yoon and Kim, Nameun and Ghosh, Mrinmoy and Lee, Sang Baek and Mok, Young Sun and Kim, Seong Bong and others, undefined
Journal: Scientific Reports (2018): 8761
Authors: Heger, Zbynek and Polanska, Hana and Krizkova, Sona and Balvan, Jan and Raudenska, Martina and Dostalova, Simona and Moulick, Amitava and Masarik, Michal and Adam, Vojtech
Journal: Colloids and Surfaces B: Biointerfaces (2017): 131--140
Authors: Lv, Xin and Song, Dong-mei and Niu, Ying-hao and Wang, Bao-shan
Journal: Apoptosis (2016): 489--501
Authors: Zou, Qianli and Zhao, Hongyou and Zhao, Yuxia and Fang, Yanyan and Chen, Defu and Ren, Jie and Wang, Xiaopu and Wang, Ying and Gu, Ying and Wu, Feipeng
Journal: Journal of medicinal chemistry (2015): 7949--7958
Authors: Kato, Hisashi and Tanaka, Goki and Masuda, Shinya and Ogasawara, Junetsu and Sakurai, Takuya and Kizaki, Takako and Ohno, Hideki and Izawa, Tetsuya
Journal: Journal of Pineal Research (2015): 267--275
Authors: Gonneaud, Alexis
Journal: (2014)
References
Authors: Feng J, Arriaga EA.
Journal: Electrophoresis (2008): 475
Authors: Roy SS, Hajnoczky G.
Journal: Methods (2008): 213
Authors: Swayne TC, Gay AC, Pon LA.
Journal: Methods Mol Biol (2007): 433
Authors: Martinez-Caballero S, Peixoto PM, Kinnally KW, Campo ML.
Journal: Anal Biochem (2007): 76
Authors: Duffy CF, MacCraith B, Diamond D, O'Kennedy R, Arriaga EA.
Journal: Lab Chip (2006): 1007
Authors: Elmore SP, Nishimura Y, Qian T, Herman B, Lemasters JJ.
Journal: Arch Biochem Biophys (2004): 145
Authors: Kristian T, Fiskum G.
Journal: Brain Res Brain Res Protoc (2004): 176
Authors: Fuller KM, Duffy CF, Arriaga EA.
Journal: Electrophoresis (2002): 1571
Authors: Nakayama S, Sakuyama T, Mitaku S, Ohta Y.
Journal: Biochem Biophys Res Commun (2002): 23
Authors: Dedov VN, Cox GC, Roufogalis BD.
Journal: Micron (2001): 653
Application notes
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Developing high-content assays for hepatoxicity with iPSC-derived cells