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DiTO™-3 [equivalent to TOTO®-3] *5 mM DMSO Solution*
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.
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.
Spectrum
Product family
Citations
View all 1 citations: Citation Explorer
Targeting metastasis-initiating cancer stem cells in gastric cancer with leukaemia inhibitory factor
Authors: Seeneevassen, Lornella and Zaafour, Anissa and Sifr{\'e}, Elodie and Genevois, Coralie and Nguyen, Tra Ly and Pobiedonoscew, Yasmine and Giese, Alban and Guignard, J{\'e}r{\^o}me and Tiffon, Camille and Rousseau, Benoit and others,
Journal: Cell Death Discovery (2024): 120
Authors: Seeneevassen, Lornella and Zaafour, Anissa and Sifr{\'e}, Elodie and Genevois, Coralie and Nguyen, Tra Ly and Pobiedonoscew, Yasmine and Giese, Alban and Guignard, J{\'e}r{\^o}me and Tiffon, Camille and Rousseau, Benoit and others,
Journal: Cell Death Discovery (2024): 120
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
Page updated on December 17, 2024
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