Cell Navigator® Mitochondrion Staining Kit *Orange Fluorescence*
Price | |
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 |
Excitation (nm) | 553 |
Emission (nm) | 576 |
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
UNSPSC | 12352200 |
Overview | ![]() ![]() |
Excitation (nm) 553 | Emission (nm) 576 |
Platform
Fluorescence microscope
Excitation | TRITC filter |
Emission | TRITC filter |
Recommended plate | Black wall/clear bottom |
Components
Example protocol
AT A GLANCE
Protocol summary
- Prepare cells
- Add Mitolite™ Orange working solution
- Incubate at 37°C for 30 minutes to 2 hours
- Analyze the cells under fluorescence microscope at Ex/Em = 540/590 nm (TRITC filter set)
Important notes
Thaw all the components at room temperature before starting the experiment.
PREPARATION OF WORKING SOLUTION
20 µL of 500X Mitolite™ Orange (Component A) into 10 mL of Live Cell Staining Buffer (Component B) to make Mitolite™ Orange working soluttion. Protect from light. Note: 20 µL of 500X Mitolite™ Orange (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™ Orange working solution.
- Incubate the cells in a 37°C, 5% CO2 incubator for 30 minutes to 2 hours.
- Replace Mitolite™ Orange working solution with Hanks and 20 mM Hepes buffer (HH buffer) or buffer of your choice (e.g. the buffer with growth medium at 1:1 concentration).
- Observe the cells using a fluorescence microscope with TRITC filter set (Ex/Em = 540/590 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™ Orange working solution.
- Incubate the cells in a 37°C, 5% CO2 incubator for 30 minutes to 2 hours.
- Replace Mitolite™ Orange working solution with Hanks and 20 mM Hepes buffer (HH buffer) or buffer of your choice (e.g. the buffer with growth medium at 1:1 concentration).
- Observe the cells using a fluorescence microscope with TRITC filter set (Ex/Em = 540/590 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
![Fluorescence images of HeLa cells stained with Cell Navigator® Mitochondrion Staining Kit *Orange Fluorescence* using fluorescence microscope with a TRITC filter set. Live cells were stained with mitochondria dye MitoLite™ Orange (Red) and imaged. After fixation, the cells were labeled with F-actin dye iFluor® 488-Phalloidin (Cat#23115, Green) and counterstained with Nuclear Blue™ DCS1 (Cat#17548, Blue).](/_next/image?url=https%3A%2F%2Fimages.aatbio.com%2Fproducts%2Ffigures-and-data%2Fcell-navigator-mitochondrion-staining-kit-orange-fluorescence%2Ffigure-for-cell-navigator-mitochondrion-staining-kit-orange-fluorescence_oRClR.jpg&w=3840&q=75)
Citations
Authors: Kan, Shiyi and Pi, Caixia and Zhang, Li and Guo, Daimo and Niu, Zhixing and Liu, Yang and Duan, Mengmeng and Pu, Xiahua and Bai, Mingru and Zhou, Chenchen and others,
Journal: Cell Communication and Signaling (2023): 1--17
Authors: Zhou, Chenchen and Wang, Chunli and Xu, Kang and Niu, Zhixing and Zou, Shujuan and Zhang, Demao and Qian, Zhiyong and Liao, Jinfeng and Xie, Jing
Journal: Bioactive Materials (2022)
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.
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Authors: Roy SS, Hajnoczky G.
Journal: Methods (2008): 213
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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
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FAQ
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