DiTO™-3 [equivalent to TOTO®-3] *5 mM DMSO Solution*
Ordering information
| Price | |
| Catalog Number | |
| Unit Size | |
| Quantity |
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 |
Physical properties
| Molecular weight | 1354.85 |
| Solvent | Water |
Spectral properties
| Extinction coefficient (cm -1 M -1) | 1540001 |
| Excitation (nm) | 642 |
| Emission (nm) | 661 |
| Quantum yield | 0.061 |
Storage, safety and handling
| H-phrase | H303, H313, H340 |
| Hazard symbol | T |
| Intended use | Research Use Only (RUO) |
| R-phrase | R20, R21, R68 |
| Storage | Freeze (< -15 °C); Minimize light exposure |
| UNSPSC | 41116134 |
| Overview |  SDSProtocol |
See also: Nucleus, Cell Viability Assays
Molecular weight 1354.85 | Extinction coefficient (cm -1 M -1) 1540001 | Excitation (nm) 642 | Emission (nm) 661 | Quantum yield 0.061 |
DiTO™-3 is chemically equivalent to TOTO®-3 (TOTO® is the trademark of Invitrogen). DiTO™-3 is a carbocyanine dimer with far-red fluorescence similar to Cy® 5 dyes. It is cell-impermeant and easily distinguished from fluorescein and rhodamine as a nuclear counterstain and dead cell indicator. It is non-fluorescent in the absence of nucleic acids, and generates a very bright fluorescence signal upon binding to DNA. DiTO™-3 gives strong and selective nuclear staining in cultured cells and in paraffin sections. Simultaneous labeling with cell-permeable Nuclear Green™ LCS1 dye and cell-impermeant DiTO®-3 can be used to assess cell viability. DiTO™-3 and Nuclear Green™ both have much greater extinction coefficients than that of DNA-bound propidium iodide.
Platform
Fluorescence microscope
| Excitation | Cy5 filter set |
| Emission | Cy5 filter set |
| Recommended plate | Black wall/clear bottom |
| Instrument specification(s) | Cy5 filter set |
Example protocol
PREPARATION OF WORKING SOLUTION
DiTO™-3 working solution
Make DiTO™-3 working solution in Hanks with 20 mM Hepes buffer (HH buffer) or buffer of your choice at your desired concentration.Note In initial experiments, it may be best to try several dye concentrations to determine the optimal concentration that yields the desired result. High dye concentration tends to cause nonspecific staining of other cellular structures.
SAMPLE EXPERIMENTAL PROTOCOL
Caution: The following protocol can be adapted for most cell types. Growth medium, cell density, the presence of other cell types and factors may influence staining. Residual detergent on glassware may also affect staining of many organisms, and cause brightly stained material to appear in solutions with or without cells present.
- Grow and treat cells as desired.
- Remove the cell culture medium and fix cells.
- Add DiTO™-3 working solution (1 to 10 µM) into the cells (either suspension or adherent cells), and stain the cells for 15 to 60 minutes.
- Remove the dye working solution and add HH buffer or buffer of your choice.
- Analyze the cellular staining with a fluorescence microscope using Cy5 filter.
Calculators
Common stock solution preparation
Table 1. Volume of Water needed to reconstitute specific mass of DiTO™-3 [equivalent to TOTO®-3] *5 mM DMSO Solution* to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.
| 0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
| 1 mM | 73.809 µL | 369.045 µL | 738.089 µL | 3.69 mL | 7.381 mL |
| 5 mM | 14.762 µL | 73.809 µL | 147.618 µL | 738.089 µL | 1.476 mL |
| 10 mM | 7.381 µL | 36.904 µL | 73.809 µL | 369.045 µL | 738.089 µL |
Molarity calculator
Enter any two values (mass, volume, concentration) to calculate the third.
| Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
| / | = | x | = |
Spectrum
Open in Advanced Spectrum Viewer
Spectral properties
| Extinction coefficient (cm -1 M -1) | 1540001 |
| Excitation (nm) | 642 |
| Emission (nm) | 661 |
| Quantum yield | 0.061 |
Product Family
Images
Figure 1. Chemical structure for DiTO™-3 [equivalent to TOTO®-3] *5 mM DMSO Solution*
References
View all 27 references: Citation Explorer
Chromatin staining of Drosophila testes
Authors: Bonaccorsi S, Giansanti MG, Cenci G, Gatti M.
Journal: Cold Spring Harb Protoc (2012)
Authors: Bonaccorsi S, Giansanti MG, Cenci G, Gatti M.
Journal: Cold Spring Harb Protoc (2012)
In vivo cancer imaging by poly(ethylene glycol)-b-poly(varepsilon-caprolactone) micelles containing a near-infrared probe
Authors: Cho H, Indig GL, Weichert J, Shin HC, Kwon GS.
Journal: Nanomedicine (2012): 228
Authors: Cho H, Indig GL, Weichert J, Shin HC, Kwon GS.
Journal: Nanomedicine (2012): 228
The fluorescent dyes TO-PRO-3 and TOTO-3 iodide allow detection of microbial cells in soil samples without interference from background fluorescence
Authors: Henneberger R, Birch D, Bergquist P, Walter M, Anitori RP.
Journal: Biotechniques (2011): 190
Authors: Henneberger R, Birch D, Bergquist P, Walter M, Anitori RP.
Journal: Biotechniques (2011): 190
Effect of Mg ions on efficiency of gene electrotransfer and on cell electropermeabilization
Authors: Haberl S, Miklavcic D, Pavlin M.
Journal: Bioelectrochemistry (2010): 265
Authors: Haberl S, Miklavcic D, Pavlin M.
Journal: Bioelectrochemistry (2010): 265
Visualizing nuclei in skin cryosections: viable options to 4'6-diamidino-2-phenylindol for confocal laser microscopy
Authors: Glaser K, Wilke K, Wepf R, Biel SS.
Journal: Skin Res Technol (2008): 324
Authors: Glaser K, Wilke K, Wepf R, Biel SS.
Journal: Skin Res Technol (2008): 324
Highly chlorinated Escherichia coli cannot be stained by propidium iodide
Authors: Phe MH, Dossot M, Guilloteau H, Block JC.
Journal: Can J Microbiol (2007): 664
Authors: Phe MH, Dossot M, Guilloteau H, Block JC.
Journal: Can J Microbiol (2007): 664
DNA labeling in living cells
Authors: Martin RM, Leonhardt H, Cardoso MC.
Journal: Cytometry A (2005): 45
Authors: Martin RM, Leonhardt H, Cardoso MC.
Journal: Cytometry A (2005): 45
Quinolinium, 1,1'-[1,3-propanediylbis[(dimethyliminio)-3,1-propanediyl]]bis[4-[3-(3-methyl-2(3 H)-benzothiazolylidene)-1-propen-1-yl]-,iodide (1:4)
Authors: Shan L., undefined
Journal: In: Molecular Imaging and Contrast Agent Database (MICAD), Bethesda (MD). (2004)
Authors: Shan L., undefined
Journal: In: Molecular Imaging and Contrast Agent Database (MICAD), Bethesda (MD). (2004)
Chlorination effect on the fluorescence of nucleic acid staining dyes
Authors: Phe MH, Dossot M, Block JC.
Journal: Water Res (2004): 3729
Authors: Phe MH, Dossot M, Block JC.
Journal: Water Res (2004): 3729
Sensitive and reliable JC-1 and TOTO-3 double staining to assess mitochondrial transmembrane potential and plasma membrane integrity: interest for cell death investigations
Authors: Zuliani T, Duval R, Jayat C, Schnebert S, Andre P, Dumas M, Ratinaud MH.
Journal: Cytometry A (2003): 100
Authors: Zuliani T, Duval R, Jayat C, Schnebert S, Andre P, Dumas M, Ratinaud MH.
Journal: Cytometry A (2003): 100