Cell Navigator® Lysosome Staining Kit *Green Fluorescence with 405 nm Excitation*
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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 |
H-phrase | H303, H313, H333 |
Hazard symbol | XN |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R22 |
UNSPSC | 12352200 |
Overview | ![]() ![]() |
Platform
Fluorescence microscope
Excitation | 405 nm |
Emission | 480 nm |
Recommended plate | Black wall/clear bottom |
Instrument specification(s) | Violet filter set |
Components
Example protocol
AT A GLANCE
Protocol summary
- Prepare cells
- Add LysoBrite™ VLG26 working solution
- Incubate at 37°C for 30 minutes to 2 hours
- Analyze the cells under fluorescence microscope at Ex/Em = 405/480 nm (Violet 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™ VLG26 stock solution (Component A) to 10 mL of Live Cell Staining Buffer (Component B) and mix well to make LysoBrite™ VLG26 working solution. Protect from light. Note: 20 µL of 500X LysoBrite™ VLG26 stock solution (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 coverslips inside a petri dish filled with the appropriate culture medium. When cells reach the desired confluence, add equal volume (such as 100 µL/well/96-well plate) of LysoBrite™ VLG26 working solution.
- Incubate the cells in a 37°C, 5% CO2 incubator for 30 minutes to 2 hours.
- Observe the cells using a fluorescence microscope with Violet filter set (Ex/Em = 405/480 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:
- Centrifuge the cells at 1,000 rpm for 5 minutes to obtain a cell pellet and aspirate the supernatant.
- Resuspend the cell pellet gently in pre-warmed growth medium, and then add equal volume of LysoBrite™ VLG26 working solution.
- Incubate the cells in a 37°C, 5% CO2 incubator for 30 minutes to 2 hours.
- Observe the cells using a fluorescence microscope with Violet filter set (Ex/Em = 405/480 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 coverslips that have been treated with BD Cell-Tak® (BD Biosciences) and stained as adherent cells.
Product Family
Images
![Image of U2OS cells stained with Cell Navigator® Lysosomal Staining Kit in a Costar black wall/clear bottom 96-well plate.](/_next/image?url=https%3A%2F%2Fimages.aatbio.com%2Fproducts%2Ffigures-and-data%2Fcell-navigator-lysosome-staining-kit-green-fluorescence-with-405-nm-excitation%2Ffigure-for-cell-navigator-lysosome-staining-kit-green-fluorescence-with-405-nm-excitation_Zoq1O.jpg&w=3840&q=75)
![To determine whether genistein or PP2 alters the distribution of the internalized BmCPV particles, we used confocal microscopy to locate internalized BmCPV virions and lysosomes. Genistein and PP2 route BmCPV to the wrong destination- lysosomes. (a) Normal BmN cells (control) were incubated with A546-labeled BmCPV virions and a lysosomal staining agent for 3 h; (b) BmN cells were treated with PP2 (0.16 μM) for 30 min, followed by incubation with A546-labeled BmCPV virions and a lysosomal staining agent for 3 h; (c) BmN cells were treated with genistein (50 μg/mL) for 1 h, followed by incubation with A546-labeled BmCPV virions and a lysosomal staining agent for 3 h; (d) quantification of the spectral overlap of BmCPV particles with lysosomes. The spectral overlap is presented as the percent co-localization (n = 21 cells). Error bars indicate standard deviations. ***P < 0.001. Source: <strong>Clathrin-mediated endocytosis is a candidate entry sorting mechanism for Bombyx mori cypovirus </strong>by Chen et al., <em>Scientific Reports, </em>May 2018.](/_next/image?url=https%3A%2F%2Fimages.aatbio.com%2Fproducts%2Ffigures-and-data%2Fcell-navigator-lysosome-staining-kit-green-fluorescence-with-405-nm-excitation%2Ffigure-for-cell-navigator-lysosome-staining-kit-green-fluorescence-with-405-nm-excitation_FFDw6.jpg&w=3840&q=75)
![Colocalization of BAPCs with dsRNA in lysosomes. 2.5 μg of dsRNA was complexed with 50 μM of BAPCs and incubated with Sf9 cells for 1 h. Lysosomes were stained with Cell Navigator. Confocal microscopy was used to check for colocalization of complexes in lysosomes. (A) Schematic representation of endocytic pathways and endosome maturation process. (B) Rh–BAPCs (red), (C) lysosomes (green), (D) bright field, and (E) merge image showing colocalization of BAPCs and the lysosomes (yellow). Source: <b>Insight into Cellular Uptake and Transcytosis of Peptide Nanoparticles in Spodoptera frugiperda Cells and Isolated Midgut</b> by McGraw <em>et. al.</em>, <em>ACS Omega</em>, March 2022.](/_next/image?url=https%3A%2F%2Fimages.aatbio.com%2Fproducts%2Ffigures-and-data%2Fcell-navigator-lysosome-staining-kit-green-fluorescence-with-405-nm-excitation%2Ffigure-for-cell-navigator-lysosome-staining-kit-green-fluorescence-with-405-nm-excitation_2bRB0.png&w=3840&q=75)
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: McGraw, Erin and Roberts, Jonathan D and Kunte, Nitish and Westerfield, Matthew and Streety, Xavier and Held, David and Avila, L Adriana
Journal: ACS omega (2022): 10933--10943
Authors: Karlsson, Johan and Tzeng, Stephany Y and Hemmati, Shayan and Luly, Kathryn M and Choi, Olivia and Rui, Yuan and Wilson, David R and Kozielski, Kristen L and Qui{\~n}ones-Hinojosa, Alfredo and Green, Jordan J
Journal: Advanced Functional Materials (2021): 2009768
Authors: Takahashi, Nobuyasu and Aoyama, Fumiyo and Sawaguchi, Akira
Journal: Microscopy (2020)
Authors: Tsai, Evaline S and Joud, Fadwa and Wiesholler, Lisa M and Hirsch, Thomas and Hall, Elizabeth AH
Journal: Analytical and Bioanalytical Chemistry (2020): 1--15
Authors: Phonesouk, Erick and Lechevallier, Séverine and Ferr, undefined and , Audrey and Rols, Marie-Pierre and Bezombes, Christine and Verelst, Marc and Golzio, Muriel
Journal: Materials (2019): 179
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: Chen, Fei and Zhu, Liyuan and Zhang, Yiling and Kumar, Dhiraj and Cao, Guangli and Hu, Xiaolong and Liang, Zi and Kuang, Sulan and Xue, Renyu and Gong, Chengliang
Journal: Scientific reports (2018): 7268
Authors: Moriwaki, Masaya and Iwamoto, Kazumasa and Niitsu, Yoshie and Matsushima, Ayako and Yanase, Yuhki and Hisatsune, Junzo and Sugai, Motoyuki and Hide, Michihiro
Journal: Allergy (2018)
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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