Cell Navigator® Lysosome Staining Kit *NIR 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) | 636 |
Emission (nm) | 651 |
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
Storage | Freeze (< -15 °C); Minimize light exposure |
UNSPSC | 12352200 |
Overview | ![]() ![]() |
Excitation (nm) 636 | Emission (nm) 651 |
Platform
Fluorescence microscope
Excitation | 630 nm |
Emission | 650 nm |
Recommended plate | Black wall/clear bottom |
Instrument specification(s) | Cy5 filter set |
Components
Example protocol
AT A GLANCE
Protocol summary
- Prepare cells
- Add LysoBrite™ NIR working solution
- Incubate at 37°C for 30 minutes
- Wash the cells
- Analyze the cells under fluorescence microscope at Ex/Em = 630/650 nm (Cy5 filter set)
Important notes
Thaw all the kit components at room temperature before starting the experiment.
PREPARATION OF WORKING SOLUTION
Add 20 µL of 500X LysoBrite™ NIR stock solution (Component A) to 10 mL of Live Cell Staining Buffer (Component B) to make LysoBrite™ NIR working solution. Protect from light. Note: 20 µL of 500X LysoBrite™ NIR (Component A) is enough for one 96-well plate. The optimal concentration of the fluorescent lysosome 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 LysoBrite™ NIR working solution.
- Incubate the cells in a 37°C, 5% CO2 incubator for 30 minutes.
- Wash the cells twice with pre-warmed (37°C) Hanks and 20 mM Hepes buffer (HBSS) or buffer of your choice, then fill the cell wells with HBSS or growth medium.
- Observe the cells using a fluorescence microscope with Cy5 filter set (Ex/Em = 630/650 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.
For suspension cells:
- Add equal volume of LysoBrite™ NIR working solution into the cells.
- Incubate the cells in a 37°C, 5% CO2 incubator for 30 minutes.
- Wash the cells twice with pre-warmed (37°C) Hanks and 20 mM Hepes buffer (HBSS) or buffer of your choice, then fill the cell wells with HBSS or growth medium.
- Observe the cells using a fluorescence microscope with Cy5 filter set (Ex/Em = 630/650 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.
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Citations
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Journal: European Biophysics Journal (2016): 1--12
Authors: Paris, Juan L and de la Torre, Paz and Manzano, Miguel and Cabanas, M Victoria and Flores, Ana I and Vallet-Regí, María
Journal: Acta biomaterialia (2016): 275--282
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Journal: Analytical Sciences (2015): 315--320
Authors: Tsai, Ching-Wei and Hu, Wei-Wen and Liu, Chih-I and Ruaan, Ruoh-Chyu and Tsai, Bing-Chang and Jin, Shiow-Lian Catherine and Chang, Yung and Chen, Wen-Yih
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