Hoechst 33258 *CAS 23491-45-4*

Hoechst 33258

CAS 23491-45-4

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.

Protocol

COO

COA

Catalog	Size	Price	Quantity
17520	100 mg	Price
17523	1 g	Price

Calculators

Common stock solution preparation

Table 1. Volume of Water needed to reconstitute specific mass of Hoechst 33258 *CAS 23491-45-4* 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	187.308 µL	936.54 µL	1.873 mL	9.365 mL	18.731 mL
5 mM	37.462 µL	187.308 µL	374.616 µL	1.873 mL	3.746 mL
10 mM	18.731 µL	93.654 µL	187.308 µL	936.54 µL	1.873 mL

Molarity calculator

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

Mass (Calculate)		Molecular weight		Volume (Calculate)		Concentration (Calculate)		Moles
mg	÷	533.88 g/mol	=	mL	x	mM	=	mmol

Citations

View all 3 citations: Citation Explorer

Protective Effects of Crataegus pinnatifida Extracts and Crataegolic Acid Against Neurotoxicity of Paraquat in PC12 Cells.

Authors:

Chang, Chi-Sen and Shen, Yuh-Chiang and Juan, Chi-Wen and Chang, Chia-Lin and Lin, Po-Kai

Journal:

Current Topics in Nutraceutical Research (2022)

3'-Oxo-tabernaelegantine A (OTNA) selectively relaxes pulmonary arteries by inhibiting AhR

Authors:

Long, Pei and Li, Yong and Wen, Qing and Huang, Maohua and Li, Songtao and Lin, Yuning and Huang, Xiaojn and chen, Minfeng and Ouyang, Jie and Ao, Yunlin and others,

Journal:

Phytomedicine (2021)

Plasma proteomic analysis of autoimmune hepatitis in an improved AIH mouse model

Authors:

Wang, Han and Yan, Wei and Feng, Zuohua and Gao, Yuan and Zhang, Liu and Feng, Xinxia and Tian, Dean

Journal:

Journal of Translational Medicine (2020): 1--16

References

View all 42 references: Citation Explorer

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

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

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

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

Physical properties

Molecular weight	533.88
Solvent	Water

Spectral properties

Extinction coefficient (cm ^-1 M ^-1)	46000 ¹
Excitation (nm)	352
Emission (nm)	454
Quantum yield	0.0340 ¹

Storage, safety and handling

Certificate of Origin	Download PDF
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
CAS	23491-45-4

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Page updated on October 11, 2025