LysoBrite™ Red DND-99
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 |
Molecular weight | 399.25 |
Solvent | DMSO |
Excitation (nm) | 573 |
Emission (nm) | 592 |
Certificate of Origin | Download PDF |
Intended use | Research Use Only (RUO) |
Storage | Freeze (< -15 °C); Minimize light exposure |
Overview | SDSProtocol |
Molecular weight 399.25 | Excitation (nm) 573 | Emission (nm) 592 |
Platform
Flow cytometer
Excitation | 532/561 nm laser |
Emission | 585/40 nm filter |
Fluorescence microscope
Excitation | TRITC filter set |
Emission | TRITC 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™ Red DND-99 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™ Red DND-99 dye to room temperature.
Dilute 20 µL of 500X LysoBrite™ Red DND-99 with 10 mL of Hanks and 20 mM HEPES buffer (HBSS) or a buffer of your choice.
Note: 20 µL of LysoBrite™ Red DND-99 dye is enough for one 96-well plate. Aliquot and store unused LysoBrite™ dye stock solutions at < -15 °C. Protect it from light and 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 | 250.47 µL | 1.252 mL | 2.505 mL | 12.523 mL | 25.047 mL |
5 mM | 50.094 µL | 250.47 µL | 500.939 µL | 2.505 mL | 5.009 mL |
10 mM | 25.047 µL | 125.235 µL | 250.47 µL | 1.252 mL | 2.505 mL |
Molarity calculator
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
/ | = | x | = |
Citations
Authors: Schmitt, M. V., Lienau, P., Fricker, G., Reichel, A.
Journal: Drug Metab Dispos (2019): 49-57
Authors: Nguyen, N. T., Park, R. M., Kim, Y. H., Min, J.
Journal: J Biotechnol (2018): 1-7
Authors: Fontana, J. M., Yin, H., Chen, Y., Florez, R., Brismar, H., Fu, Y.
Journal: Int J Nanomedicine (2017): 8615-8629
Authors: Pereira, L. C., Duarte, F. V., Varela, A. T., Rolo, A. P., Palmeira, C. M., Dorta, D. J.
Journal: Toxicology (2017): 59-65
Authors: Balashova, N., Dhingra, A., Boesze-Battaglia, K., Lally, E. T.
Journal: Mol Oral Microbiol (2016): 106-14
Authors: Zhu, S., Rea, S. L., Cheng, T., Feng, H. T., Walsh, J. P., Ratajczak, T., Tickner, J., Pavlos, N., Xu, H. Z., Xu, J.
Journal: J Cell Biochem (2016): 1464-70
Authors: Majzoub, R. N., Wonder, E., Ewert, K. K., Kotamraju, V. R., Teesalu, T., Safinya, C. R.
Journal: J Phys Chem B (2016): 6439-53
Authors: Mallo, N., Lamas, J., Defelipe, A. P., Decastro, M. E., Sueiro, R. A., Leiro, J. M.
Journal: Parasitology (2016): 576-87
Authors: Harlan, F. K., Lusk, J. S., Mohr, B. M., Guzikowski, A. P., Batchelor, R. H., Jiang, Y., Naleway, J. J.
Journal: PLoS One (2016): e0156312
Authors: Schiessl, I. M., Hammer, A., Kattler, V., Gess, B., Theilig, F., Witzgall, R., Castrop, H.
Journal: J Am Soc Nephrol (2016): 731-44
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?