Cell Navigator® Lysosome Staining Kit *Deep Red 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) | 597 |
Emission (nm) | 619 |
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
UNSPSC | 12352200 |
Overview | ![]() ![]() |
Excitation (nm) 597 | Emission (nm) 619 |
Platform
Fluorescence microscope
Excitation | Texas Red filter |
Emission | Texas Red filter |
Recommended plate | Black wall/clear bottom |
Components
Example protocol
AT A GLANCE
Protocol summary
- Prepare cells
- Add Lysolite™ Deep Red working solution
- Incubate at 37°C for 30 - 60 minutes
- Add Staining Buffer B
- Incubate at room temperature for 15 - 30 minutes
- Analyze the cells under fluorescence microscope at Ex/Em = 590/620 nm (Texas Red filter set)
Important notes
Thaw all the kit components at room temperature before starting the experiment.
PREPARATION OF WORKING SOLUTION
Add 10 µL of 500X Lysolite™ Deep Red (Component A) into 5 mL of Live Cell Staining Buffer A (Component B) and mix well to make Lysolite™ Deep Red working solution. Protect from light. Note: 10 µL of 500X Lysolite™ Deep Red (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 1/2 volume (such as 50 µL/well/96-well plate) of Lysolite™ Deep Red working solution.
- Incubate the cells in a 37°C, 5% CO2 incubator for 30 to 60 minutes.
- Add 50 µL/well (96-well plate) of Staining Buffer B (Component C) into Lysolite™ Deep Red working solution plate.
- Incubate at room temperature for 15 - 30 minutes.
- Observe the cells using a fluorescence microscope with Texas Red filter set (Ex/Em = 590/620 nm). Note: The LysoBrite™ Deep Red has minimal or no cell toxicity; it can be used for cell tracking. For cell tracking purpose, skip adding Staining Buffer B: Simply replace the dye-working solution with growth medium, and then observe under fluorescence microscope. 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:
- Centrifuge the cells at 1000 rpm for 5 minutes to obtain a cell pellet and aspirate the supernatant.
- Resuspend the cell pellet gently in pre-warmed growth medium (such as 100 µL/tube), and then add 1/2 volume (50 µL/tube) of Lysolite™ Deep Red working solution.
- Incubate the cells in a 37°C, 5% CO2 incubator for 30 to 60 minutes.
- Add 50 µL/tube of Staining Buffer B (Component C) into the Lysolite™ Deep Red working solution plate.
- Incubate at room temperature for 15 - 30 minutes.
- Observe the cells using a fluorescence microscope with Texas red filter set (Ex/Em = 590/620 nm). Note: The LysoBrite™ Deep Red has minimal or no cell toxicity; it can be used for cell tracking. For cell tracking purpose, skip adding Staining Buffer B: Simply replace the dye-working solution with growth medium, and then observe under fluorescence microscope. 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
Authors: Ben-Akiva, Elana and Karlsson, Johan and Hemmati, Shayan and Yu, Hongzhe and Tzeng, Stephany Y and Pardoll, Drew M and Green, Jordan J
Journal: Proceedings of the National Academy of Sciences (2023): e2301606120
Authors: Martin, Victor and Ribeiro, Isabel AC and Alves, Marta M and Gon{\c{c}}alves, L{\'\i}dia and Almeida, Ant{\'o}nio J and Grenho, Liliana and Fernandes, Maria H and Santos, Catarina F and Gomes, Pedro S and Bettencourt, Ana F
Journal: International journal of pharmaceutics (2019): 118821
Authors: Lechevallier, S{\'e}verine and Mauricot, Robert and Gros-Dagnac, H{\'e}l{\`e}ne and Chevreux, Sylviane and Lemercier, Gilles and Phonesouk, Erick and Golzio, Muriel and Verelst, Marc
Journal: ChemPlusChem (2017): 770--777
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Journal: ChemPlusChem (2017)
Authors: Wilson, David R and Routkevitch, Denis and Rui, Yuan and Mosenia, Arman and Wahlin, Karl J and Quinones-Hinojosa, Alfredo and Zack, Donald J and Green, Jordan J
Journal: Molecular Therapy (2017)
Authors: Cemma, Marija and Grinstein, Sergio and Brumell, John H
Journal: Autophagy (2016): 1440--1446
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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
Authors: Sato, Takaya and Sato, Yusuke and Iwai, Kenta and Kuge, Shusuke and Teramae, Norio and Nishizawa, Seiichi
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
Journal: International journal of pharmaceutics (2015): 498--505
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