Cell Navigator® Lysosome Staining Kit *Orange Fluorescence*
Ordering information
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Additional ordering information
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 |
Spectral properties
Excitation (nm) | 543 |
Emission (nm) | 565 |
Storage, safety and handling
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
UNSPSC | 12352200 |
Alternative formats
Related products
Overview | SDSProtocol |
See also: Lysosomes, Mitochondria
Excitation (nm) 543 | Emission (nm) 565 |
Lysosomes are cellular organelles which contain acid hydrolase enzymes to break up waste materials and cellular debris. Lysosomes digest excess or worn-out organelles, food particles, and engulfed viruses or bacteria. The membrane around a lysosome allows the digestive enzymes to work at pH 4.5. The interior of the lysosomes is acidic (pH 4.5-4.8) compared to the slightly alkaline cytosol (pH 7.2). The lysosome maintains this pH differential by pumping protons from the cytosol across the membrane via proton pumps and chloride ion channels. Our Cell Navigator® fluorescence imaging kits are a set of fluorescence imaging tools for labeling sub-cellular organelles such as membranes, lysosomes, mitochondria, nuclei, etc. The selective labeling of live cell compartments provides a powerful method for studying cellular events in a spatial and temporal context. This particular kit is designed to label lysosomes of live cells in orange fluorescence. LysoBrite™ Orange, the proprietary lysotropic dye used in the kit, selectively accumulates in lysosomes probably via the lysosome pH gradient. The lysotropic indicator is a hydrophobic compound that easily permeates intact live cells, and trapped in lysosomes after it gets into cells. Its fluorescence is significantly enhanced upon entering lysosomes. This key feature significantly reduces its staining background and makes it useful for a variety of studies, including cell adhesion, chemotaxis, multidrug resistance, cell viability, apoptosis and cytotoxicity. It is suitable for proliferating and non-proliferating cells, and can be used for both suspension and adherent cells. LysoBrite™ dyes significantly outperform the equivalent LysoTracker ™dyes (from Invitrogen). LysoBrite™ dyes can stay in live cells for more than a week with very minimal cell toxicity while the LysoTracker dyes can only be used for a few hours. LysoBrite™ dyes can survive a few generations of cell division. In addition, LysoBrite™ dyes are much more photostable than the LysoTracker dyes.
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 LysoBrite™ Orange working solution
- Incubate at 37°C for 30 minutes
- Wash the cells
- Analyze the cells under fluorescence microscope at Ex/Em = 540/570 nm (TRITC filter set)
Important notes
Thaw all the kit components at room temperature before starting the experiment.
PREPARATION OF WORKING SOLUTION
20 µL of 500X LysoBrite™ Orange (Component A) to 10 mL of Live Cell Staining Buffer (Component B) to make LysoBrite™ Orange working solution. Note: 20 µL of 500X LysoBrite™ Orange (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™ Orange 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, fill the cell wells with HBSS or growth medium.
- Observe the cells using a fluorescence microscope with TRITC filter set (Ex/Em = 540/570 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™ Orange 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, fill the cell wells with HBSS or growth medium.
- Observe the cells using a fluorescence microscope with TRITC filter set (Ex/Em = 540/570 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
View all 14 citations: Citation Explorer
Biodegradable lipophilic polymeric mRNA nanoparticles for ligand-free targeting of splenic dendritic cells for cancer vaccination
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: 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
Understanding intracellular trafficking and anti-inflammatory effects of minocycline chitosan-nanoparticles in human gingival fibroblasts for periodontal disease treatment
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: 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
Silica-Based Nanoparticles as Bifunctional and Bimodal Imaging Contrast Agents
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
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
Silica-based nanoparticles as bi-functional and bi-modal imaging contrast agents
Authors: Lechevallier, Séverine and Mauricot, Robert and Gros-Dagnac, Hélène and Chevreux, Sylviane and Lemercier, Gilles and Phonesouk, Erick and Golzio, Muriel and Verelst, Marc
Journal: ChemPlusChem (2017)
Authors: Lechevallier, Séverine and Mauricot, Robert and Gros-Dagnac, Hélène and Chevreux, Sylviane and Lemercier, Gilles and Phonesouk, Erick and Golzio, Muriel and Verelst, Marc
Journal: ChemPlusChem (2017)
A Triple-Fluorophore Labeled Nucleic Acid pH Nanosensor to Investigate Non-Viral Gene Delivery
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: 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)
Rhodamine bound maghemite as a long-term dual imaging nanoprobe of adipose tissue-derived mesenchymal stromal cells
Authors: Cmiel, Vratislav and Skopalik, Josef and Polakova, Katerina and Solar, Jan and Havrdova, Marketa and Milde, David and Justan, Ivan and Magro, {cmiel2016rhodamine, title={Rhodamine bound maghemite as a long-term dual imaging nanoprobe of adipose tissue-derived mesenchymal stromal cells
Journal: European Biophysics Journal (2016): 1--12
Authors: Cmiel, Vratislav and Skopalik, Josef and Polakova, Katerina and Solar, Jan and Havrdova, Marketa and Milde, David and Justan, Ivan and Magro, {cmiel2016rhodamine, title={Rhodamine bound maghemite as a long-term dual imaging nanoprobe of adipose tissue-derived mesenchymal stromal cells
Journal: European Biophysics Journal (2016): 1--12
Decidua-derived mesenchymal stem cells as carriers of mesoporous silica nanoparticles. In vitro and in vivo evaluation on mammary tumors
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: 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
Fluorescence imaging of siRNA delivery by peptide nucleic acid-based probe
Authors: Sato, Takaya and Sato, Yusuke and Iwai, Kenta and Kuge, Shusuke and Teramae, Norio and Nishizawa, Seiichi
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The consideration of indolicidin modification to balance its hemocompatibility and delivery efficiency
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
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
Endocytosed β2-microglobulin amyloid fibrils induce necrosis and apoptosis of rabbit synovial fibroblasts by disrupting endosomal/lysosomal membranes: a novel mechanism on the cytotoxicity of amyloid fibrils
Authors: Okoshi, Tadakazu and Yamaguchi, Itaru and Ozawa, Daisaku and Hasegawa, Kazuhiro and Naiki, Hironobu
Journal: PloS one (2015): e0139330
Authors: Okoshi, Tadakazu and Yamaguchi, Itaru and Ozawa, Daisaku and Hasegawa, Kazuhiro and Naiki, Hironobu
Journal: PloS one (2015): e0139330
References
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Lectin-histochemical and -cytochemical study of periodic acid Schiff-positive lysosome granules as a histological feature of the female mouse kidney
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Journal: Histol Histopathol (2002): 1017
Authors: Yabuki A, Suzuki S, Matsumoto M, Nishinakagawa H.
Journal: Histol Histopathol (2002): 1017
Alz-50/Gallyas-positive lysosome-like intraneuronal granules in Alzheimer's disease and control brains
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Journal: Neurosci Lett (1998): 113
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Application notes
A Novel Fluorescent Probe for Imaging and Detecting Hydroxyl Radical in Living Cells
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
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?
Do you have any fixable mitochondria staining assay kits?
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?
Do you have any fixable mitochondria staining assay kits?