LysoBrite™ NIR
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Catalog Number | |
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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 |
Molecular weight | 730.89 |
Solvent | DMSO |
Excitation (nm) | 636 |
Emission (nm) | 651 |
Certificate of Origin | Download PDF |
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 |
LysoBrite™ Blue |
LysoBrite™ Green |
LysoBrite™ Orange |
LysoBrite™ Red |
LysoBrite™ Deep Red |
LysoBrite™ Red DND-99 |
LysoBrite™ Green DND-26 |
Overview | SDSProtocol |
Molecular weight 730.89 | Excitation (nm) 636 | Emission (nm) 651 |
Platform
Flow cytometer
Excitation | 640 nm laser |
Emission | 660/20 nm filter |
Instrument specification(s) | APC channel |
Fluorescence microscope
Excitation | Cy5 filter set |
Emission | Cy5 filter set |
Recommended plate | Black wall/clear bottom |
Example protocol
AT A GLANCE
Prepare cells.
Add dye working solution.
Incubate at 37 °C for 30 minutes.
Wash the cells.
Analyze under a fluorescence microscope.
The LysoBrite™ NIR stock solution provided is 500X in DMSO. It should be stable for at least 6 months if stored at -20 °C and protected from light. Avoid freeze/thaw cycles.
PREPARATION OF WORKING SOLUTION
Warm LysoBrite™ NIR dye to room temperature.
Dilute 20 µL of 500X LysoBrite™ NIR with 10 mL of Hanks and 20 mM HEPES buffer (HBSS) or buffer of your choice.
Note: 20 µL of LysoBrite™ NIR dye is enough for one 96-well plate. Aliquot and store any unused LysoBrite™ dye stock solution at < -15 °C, protected from light. Avoid repeated freeze-thaw cycles.
Note: 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.
SAMPLE EXPERIMENTAL PROTOCOL
This protocol only provides a guideline and should be modified according to your specific needs.
Grow cells 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 an equal volume of the dye-working solution (from Preparation of Working Solution Step 2).
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 fitted with the desired filter set.
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.
Add an equal volume of the dye-working solution (from Preparation of Working Solution Step 2).
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 fitted with the desired filter set.
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.
Note: Suspension cells may be attached to coverslips treated with BD Cell-Tak® (BD Biosciences) and stained as adherent cells (see Protocol for Preparing and Staining Adherent Cells).
Calculators
Common stock solution preparation
0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
1 mM | 136.819 µL | 684.097 µL | 1.368 mL | 6.841 mL | 13.682 mL |
5 mM | 27.364 µL | 136.819 µL | 273.639 µL | 1.368 mL | 2.736 mL |
10 mM | 13.682 µL | 68.41 µL | 136.819 µL | 684.097 µL | 1.368 mL |
Molarity calculator
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
/ | = | x | = |
Product Family
Name | Excitation (nm) | Emission (nm) |
LysoBrite™ Blue | 434 | 480 |
LysoBrite™ Green | 501 | 510 |
LysoBrite™ Orange | 543 | 565 |
LysoBrite™ Red | 576 | 596 |
LysoBrite™ Deep Red | 597 | 619 |
Images
Citations
Authors: Wehling, Arne and Loeffler, Dirk and Zhang, Yang and Kull, Tobias and Donato, Cinzia and Szczerba, Barbara and Camargo Ortega, Germ{\'a}n and Lee, Minkyoung and Moor, Andreas and G{\"o}ttgens, Bertie and others,
Journal: Blood, The Journal of the American Society of Hematology (2022): 1482--1495
Authors: Bheri, Sruti and Kassouf, Brandon P and Park, Hyun-Ji and Hoffman, Jessica R and Davis, Michael E
Journal: Journal of Cardiovascular Development and Disease (2021): 135
Authors: Xu, Mei-Qi and Hao, Yan-Li and Wang, Jing-Ru and Li, Zhuo-Yue and Li, Hui and Feng, Zhen-Han and Wang, Hui and Wang, Jing-Wen and Zhang, Xuan
Journal: International Journal of Nanomedicine (2021): 7269
Authors: Zhou, Xinyang and Pan, Yufei and Li, Zheng and Li, Huantong and Wu, Jing and Ma, Yuan and Guan, Zhu and Yang, Zhenjun
Journal: ACS Applied Bio Materials (2020): 6297--6309
Authors: Xue, Legang and Liu, Pei
Journal: Biochemical and Biophysical Research Communications (2020)
Authors: Zhang, Shuang and Li, Zhan-Tao and Liu, Man and Wang, Jing-Ru and Xu, Mei-Qi and Li, Zhuo-Yue and Duan, Xiao-Chuan and Hao, Yan-Li and Zheng, Xiu-Chai and Li, Hui and others, undefined
Journal: Journal of Controlled Release (2018)
Authors: Cemma, Marija and Grinstein, Sergio and Brumell, John H
Journal: Autophagy (2016): 1440--1446
Authors: Myung, Ja Hye and Gajjar, Khyati A and Chen, Jihua and Molokie, Robert E and Hong, Seungpyo
Journal: Analytical chemistry (2014): 6088--6094
Authors: Lei, Bo and Chen, Xiaofeng and Han, Xue and Zhou, Jiaan
Journal: Journal of Materials Chemistry (2012): 16906--16913
Authors: Fedotov, Sergei and Waigh, Thomas A
References
Authors: Gough AH, Johnston PA.
Journal: Methods Mol Biol (2007): 41
Authors: Hoffman AF, Garippa RJ.
Journal: Methods Mol Biol (2007): 19
Authors: Giuliano KA., undefined
Journal: Methods Mol Biol (2007): 189
Authors: Taylor DL., undefined
Journal: Methods Mol Biol (2007): 3
Authors: Martinez ED, Dull AB, Beutler JA, Hager GL.
Journal: Methods Enzymol (2006): 21
Authors: Bowen WP, Wylie PG.
Journal: Assay Drug Dev Technol (2006): 209
Authors: Hudson CC, Oakley RH, Sjaastad MD, Loomis CR.
Journal: Methods Enzymol (2006): 63
Authors: Werner T, Liebisch G, Gr and l M, Schmitz G.
Journal: Cytometry A (2006): 200
Authors: Wolff M, Haasen D, Merk S, Kroner M, Maier U, Bordel S, Wiedenmann J, Nienhaus GU, Valler M, Heilker R.
Journal: Comb Chem High Throughput Screen (2006): 339
Authors: O'Brien P J, Irwin W, Diaz D, Howard-Cofield E, Krejsa CM, Slaughter MR, Gao B, Kaludercic N, Angeline A, Bernardi P, Brain P, Hougham C.
Journal: Arch Toxicol (2006): 580
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
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?