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Hoechst 33258

Hoechst 33258 is a blue to blue-green fluorescent live cell dye that can label DNA. Hoechst 33258 can specifically bind to the minor groove of DNA (and tends to bind to A/T-rich DNA), resulting in a significant increase in fluorescence intensity. Hoechst 33258 can cross the cell membrane and cause changes in DNA structure, such as G2/M phase arrest. Hoechst 33258 can bind to live or fixed cells, and the fluorescence intensity increases with increasing solution pH. As a DNA-specific probe, Hoechst 33258 can be used to detect DNA content, analyze cell cycle, etc. The excitation wavelength of Hoechst 33258 is 350-365 nm, and the emission wavelength is 460-490 nm[1][2][3].

Product Specifications

CAS Number

23491-44-3

Product Name Alternative

BisBenzimide H 33258; H 33258

UNSPSC

12171500

Hazard Statement

H302, H315, H319, H335

Target

DNA Alkylator/Crosslinker; DNA Stain; Fluorescent Dye

Type

Dye Reagents

Related Pathways

Cell Cycle/DNA Damage; Others

Applications

Cancer-programmed cell death

Field of Research

Others

Assay Protocol

https://www.medchemexpress.com/Hoechst-33258.html

Purity

99.93

Solubility

DMSO : 50 mg/mL (ultrasonic)

Smiles

OC1=CC=C(C2=NC3=CC=C(C4=NC5=CC=C(N6CCN(C)CC6)C=C5N4)C=C3N2)C=C1

Molecular Formula

C25H24N6O

Molecular Weight

424.50

Precautions

H302, H315, H319, H335

References & Citations

[1]Stokke T, et al. Multiple binding modes for Hoechst 33258 to DNA. J Histochem Cytochem. 1985 Apr;33 (4) :333-8.|[2]Wang XJ, et al. Newly synthesized bis-benzimidazole derivatives exerting anti-tumor activity through induction of apoptosis and autophagy. Bioorg Med Chem Lett. 2012 Oct 1;22 (19) :6297-300.|[3]Pjura PE, et al. Binding of Hoechst 33258 to the minor groove of B-DNA. J Mol Biol. 1987 Sep 20;197 (2) :257-71.

Shipping Conditions

Blue Ice

Storage Conditions

-20°C (Powder, protect from light)

Scientific Category

Dye Reagents

Clinical Information

No Development Reported

Citation 01

Acta Biochim Biophys Sin (Shanghai) . 2019 Aug 5;51 (8) :814-825. |Authorea. 2025 Sep 20.|Biochem Biophys Res Commun. 2020 Jun 30;527 (3) :805-810.|Biochem Biophys Res Commun. 2025 Apr 6:762:151769.|Biochem Cell Biol. 2021 Aug;99 (4) :403-413.|BIOTECHNOLOGY BULLETIN 2018, 34 (12) :172-178|Cancer Manag Res. 2020 Sep 25;12:9097-9111.|Cancer Manag Res. 2020 Sep 28;12:9197-9209.|Chem Eng J. 365 (2019) 270-281.|Discov Oncol. 2025 Feb 18;16 (1) :199.|Food Biosci. 11 December 2021, 101501.|Int Immunopharmacol. 2023 Jun:119:110136.|Int J Biochem Cell Biol. 2017 Mar:84:75-88.|J Cell Biochem. 2019 Jun;120 (6) :9181-9192.|Nat Commun. 2020 Feb 5;11 (1) :719.|Phytomedicine. 2025 Jul:142:156632.|Research Square Preprint. 2020 Jun.|Virulence. 2022 Dec;13 (1) :444-457.|Linkoping University. 2025.

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