Hoechst 33258 *20 mM solution in water*

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Chemical structure for Hoechst 33258 *20 mM solution in water*
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5 mL 17525 $95

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Ex/Em (nm)352/461
CAS #23491-45-4
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Category Nucleic Acid Detection
DNA Detection
Related Labeling Cells
Fluorescence Imaging
Secondary Reagents
The Hoechst stains are a family of fluorescent stains for labeling DNA in fluorescence microscopy. Because these fluorescent stains label DNA, they are also commonly used to visualize nuclei and mitochondria. Two of these closely related bis-benzimides are commonly used: Hoechst 33258 and Hoechst 33342. Both dyes are excited by ultraviolet light at around 350 nm, and both emit blue/cyan fluorescence light around an emission maximum at 461 nm. The Hoechst stains may be used on live or fixed cells, and are often used as a substitute for another nucleic acid stain, DAPI. The key difference between them is that the additional ethyl group of Hoechst 33342 renders it more lipophilic, and thus more able to cross intact cell membranes. In some applications, Hoechst 33258 is significantly less permeant. These dyes can also be used to detect the contents of a sample DNA by plotting a standard emission-to-content curve.

Common stock solution preparation

Table 1. Volume of Water needed to reconstitute specific mass of Hoechst 33258 *20 mM solution in water* to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

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Table 2. Enter any two values (mass, volume, concentration) to calculate the third.

Mass Molecular weight Volume Concentration Moles
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This protocol only provides a guideline, and should be modified according to your specific needs.

The following procedure can be adapted for most cell types. Growth medium, cell density, the presence of other cell types and other factors may influence staining. Residual detergent on glassware may also affect real or apparent staining of many organisms, causing brightly stained material to appear in solutions with or without cells present.

Pellet cells by centrifugation and resuspend the cells in buffered salt solutions or media, with optimal dye binding at pH 7.4. Adherent cells in culture may be stained in situ on cover slips or in the cell culture wells. Add Hoechst stain using the concentrations between 0.5 and 5 µM and incubate it for 15 to 60 minutes as a guide. In initial experiments, it may be best to try several dye concentrations over the entire suggested range to determine the concentration that yields optimal staining.

References & Citations

Amine terminated G-6 PAMAM dendrimer and its interaction with DNA probed by Hoechst 33258
Authors: Choi YS, Cho TS, Kim JM, Han SW, Kim SK.
Journal: Biophys Chem (2006): 142

Design of new bidentate ligands constructed of two Hoechst 33258 units for discrimination of the length of two A3T3 binding motifs
Authors: Tanada M, Tsujita S, Sasaki S.
Journal: J Org Chem (2006): 125

DNA sequence recognition by Hoechst 33258 conjugates of hairpin pyrrole/imidazole polyamides
Authors: Correa BJ, Canzio D, Kahane AL, Reddy PM, Bruice TC.
Journal: Bioorg Med Chem Lett (2006): 3745

Recognition of B-DNA by neomycin--Hoechst 33258 conjugates
Authors: Willis B, Arya DP.
Journal: Biochemistry (2006): 10217

The Hoechst 33258 Covalent Dimer Covers a Total Turn of the Double-stranded DNA
Authors: Streltsov SA, Gromyko AV, Oleinikov VA, Zhuze AL.
Journal: J Biomol Struct Dyn (2006): 285

Activity of Hoechst 33258 against Pneumocystis carinii f. sp. muris, Candida albicans, and Candida dubliniensis
Authors: Disney MD, Stephenson R, Wright TW, Haidaris CG, Turner DH, Gigliotti F.
Journal: Antimicrob Agents Chemother (2005): 1326

Association of the minor groove binding drug Hoechst 33258 with d(CGCGAATTCGCG)2: volumetric, calorimetric, and spectroscopic characterizations
Authors: Han F, Taulier N, Chalikian TV.
Journal: Biochemistry (2005): 9785

Hoechst 33258 as a pH-sensitive probe to study the interaction of amine oxide surfactants with DNA
Authors: Goracci L, Germani R, Savelli G, Bassani DM.
Journal: Chembiochem (2005): 197

Hydration changes in the association of Hoechst 33258 with DNA
Authors: Kiser JR, Monk RW, Smalls RL, Petty JT.
Journal: Biochemistry (2005): 16988

Recognition of a 10 base pair sequence of DNA and stereochemical control of the binding affinity of chiral hairpin polyamide-Hoechst 33258 conjugates
Authors: Reddy PM, Toporowski JW, Kahane AL, Bruice TC.
Journal: Bioorg Med Chem Lett (2005): 5531

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Additional Documents

Safety Data Sheet (SDS)

1. Nucleic Acid Detection Probes & Assay Kits

Application Notes
1. Fluorescence Microscopy: Imaging Organelles

Certificate of Analysis