Cell Navigator® Mitochondrion Staining Kit *Blue Fluorescence*
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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) | 344 |
Emission (nm) | 469 |
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
UNSPSC | 12352200 |
Overview | SDSProtocol |
Excitation (nm) 344 | Emission (nm) 469 |
Platform
Fluorescence microscope
Excitation | DAPI filter |
Emission | DAPI filter |
Recommended plate | Black wall/clear bottom |
Components
Example protocol
AT A GLANCE
Protocol summary
- Prepare cells
- Add Mitolite™ Blue working solution
- Incubate at 37°C for 30 minutes to 2 hours
- Analyze the cells under fluorescence microscope at Ex/Em = 360/445 nm (DAPI 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™ Blue (Component A) into 10 mL of Live Cell Staining Buffer (Component B) to make Mitolite™ Blue working solution. Protect from light. Note: 20 µL of 500X Mitolite™ Blue (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™ Blue working solution.
- Incubate the cells in a 37°C, 5% CO2 incubator for 30 minutes to 2 hours.
- Replace Mitolite™ Blue 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 DAPI filter set (Ex/Em = 360/445 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™ Blue working solution.
- Incubate the cells in a 37°C, 5% CO2 incubator for 30 minutes to 2 hours.
- Replace Mitolite™ Blue 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 DAPI filter set (Ex/Em = 360/445 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: 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
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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
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Journal: Brain Res Brain Res Protoc (2004): 176
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Journal: Biochem Biophys Res Commun (2002): 23
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Journal: Micron (2001): 653
Application notes
Fluorescent Oligonucleotide Labeling Reagents
Monitoring of Mitochondrial Membrane Potential Changes in Live Cells Using JC-10
Selective Analysis of RNA in Live and Fixed Cells with StrandBrite RNA Green
Cell Loading Protocol For Fluorescent pH Indicator, BCECF-AM
FAQ
I ordered your phalloidin-amine (Cat #5302) so I can conjugate it to my oligo. Do you have a recommended protocol I can use?
What dye works best for staining and tracking lysosomes in live cells for several hours?
How can I lyse my cells without lysing the nuclear membrane?
Do you have any dual-fluorescence nucleic acid stains that interact with both DNA and RNA?