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Hoechst 33342 *Ultrapure Grade*

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.

Spectrum

Product family

NameExcitation (nm)Emission (nm)Extinction coefficient (cm -1 M -1)Quantum yield
Hoechst 33258 *CAS 23491-45-4*3524544600010.03401
Hoechst 33258 *20 mM solution in water*3524544600010.03401
Hoechst 34580 *CAS 911004-45-0*371438--
Hoechst 34580 *20 mM solution in water*371438--

Citations

View all 35 citations: Citation Explorer
Cigarette smoke compromises macrophage innate sensing in response to pneumococcal infection
Authors: Liao, Wei-Chih and Chou, Chia-Huei and Ho, Mao-Wang and Chen, Jo-Tsen and Chou, Shu-Ling and Huang, Mei-Zi and Bui, Ngoc-Niem and Wu, Hui-Yu and Lee, Chi-Fan and Huang, Wei-Chien and others,
Journal: Journal of Microbiology, Immunology and Infection (2024)
LGP2 Facilitates Bacterial Escape through Binding Peptidoglycan via EEK Motif and Suppressing NOD2--RIP2 Axis in Cyprinidae and Xenocyprididae Families
Authors: Liang, Bo and Li, Wenqian and Yang, Chunrong and Su, Jianguo
Journal: The Journal of Immunology (2024)
TLR7 neo-functionalizes to sense dsRNA and trigger antiviral and antibacterial immunity in non-tetrapod vertebrates
Authors: Jiang, Rui and Zhu, Wentao and Liao, Zhiwei and Yang, Chunrong and Su, Jianguo
Journal: iScience (2023): 108315
CHST11-modified chondroitin 4-sulfate as a potential therapeutic target for glioblastoma
Authors: Lin, You-Cheng and Chu, Yin-Hung and Liao, Wen-Chieh and Chen, Chia-Hua and Hsiao, Wen-Chuan and Ho, Ying-Jui and Yang, Meng-Yin and Liu, Chiung-Hui
Journal: American Journal of Cancer Research (2023): 2998
Biofilm formation on the surface of monazite and xenotime during bioleaching
Authors: van Alin, Arya and Corbett, Melissa K and Fathollahzadeh, Homayoun and Tjiam, M Christian and Rickard, William DA and Sun, Xiao and Putnis, Andrew and Eksteen, Jacques and Kaksonen, Anna H and Watkin, Elizabeth
Journal: Microbial Biotechnology (2023)

References

View all 42 references: Citation Explorer
Usefulness of a triple fluorochrome combination Merocyanine 540/Yo-Pro 1/Hoechst 33342 in assessing membrane stability of viable frozen-thawed spermatozoa from Estonian Holstein AI bulls
Authors: Hallap T, Nagy S, Jaakma U, Johannisson A, Rodriguez-Martinez H.
Journal: Theriogenology (2006): 1122
Fatty acid synthase and its mRNA concentrations are decreased at different times following Hoechst 33342-induced apoptosis in BC3H-1 myocytes
Authors: Zhang X, Kiechle FL.
Journal: Ann Clin Lab Sci (2006): 185
Resistance mechanism development to the topoisomerase-I inhibitor Hoechst 33342 by Leishmania donovani
Authors: Marquis JF, Hardy I, Olivier M.
Journal: Parasitology (2005): 197
The DNA minor groove binding agents Hoechst 33258 and 33342 enhance recombinant adeno-associated virus (rAAV) transgene expression
Authors: Li L, Yang L, Kotin RM.
Journal: J Gene Med (2005): 420
Acid-base and electronic structure-dependent properties of Hoechst 33342
Authors: Aleman C, Namba AM, Casanovas J.
Journal: J Biomol Struct Dyn (2005): 29
Page updated on November 21, 2024

Ordering information

Price
Unit size
100 mg
1 g
Catalog Number
1753017533
Quantity
Add to cart

Additional ordering information

Telephone1-800-990-8053
Fax1-800-609-2943
Emailsales@aatbio.com
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Request quotation

Physical properties

Molecular weight

561.93

Solvent

Water

Spectral properties

Excitation (nm)

352

Emission (nm)

454

Storage, safety and handling

Certificate of OriginDownload PDF
H-phraseH303, H313, H340
Hazard symbolT
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R68

Storage

Freeze (< -15 °C); Minimize light exposure
UNSPSC41116134

CAS

875756-97-1

Platform

Fluorescence microscope

Excitation350 nm
Emission461 nm
Recommended plateBlack wall, clear bottom
HeLa cells were incubated in 1X HBSS buffer with 5% serum to induce starvation. Following starvation, cells were treated with Autophagy Green&trade; (Cat No. 23002) working solution for 20 minutes in a 37&deg;C, 5% CO<sub>2</sub>&nbsp;incubator and then washed 3 times. Nuclei were labeled with Hoechst 33342 (Cat No. 17530). Lysosomes were labeled with LysoBrite&trade; Orange (Cat No. 22657).
HeLa cells were incubated in 1X HBSS buffer with 5% serum to induce starvation. Following starvation, cells were treated with Autophagy Green&trade; (Cat No. 23002) working solution for 20 minutes in a 37&deg;C, 5% CO<sub>2</sub>&nbsp;incubator and then washed 3 times. Nuclei were labeled with Hoechst 33342 (Cat No. 17530). Lysosomes were labeled with LysoBrite&trade; Orange (Cat No. 22657).
HeLa cells were incubated in 1X HBSS buffer with 5% serum to induce starvation. Following starvation, cells were treated with Autophagy Green&trade; (Cat No. 23002) working solution for 20 minutes in a 37&deg;C, 5% CO<sub>2</sub>&nbsp;incubator and then washed 3 times. Nuclei were labeled with Hoechst 33342 (Cat No. 17530). Lysosomes were labeled with LysoBrite&trade; Orange (Cat No. 22657).
HeLa cells were seeded in 96-well microplates and incubated at 37 °C, 5% CO<sub>2</sub> for 24 hours. Cells were then stained with 5 or 10 µM Hoechst 33342 for 30 minutes at 37 °C, washed, and imaged on a Keyence BZ-X microscope. Afterward, cells were fixed with 4% formaldehyde for 20 minutes at RT, washed and imaged.
<strong>Workflow for the &ldquo;three-in-one&rdquo; cell death screening assay.&nbsp;</strong>HUVECs growing in the 96-well plate for 48 hours are exposed to NPs for 24 hours. Three types of cell death are evaluated simultaneously. A) Cell necrosis is measured spectrophotometrically after mixing an aliquot of cell supernatant with LDH substrate. B) Cell viability is assessed by adding WST-8 substrate to the cells. After three hours of incubation, aliquots of the reaction mixture are transferred into the new plate and measured spectrophotometrically. C) Cell apoptosis is detected after incubating the cells with Hoechst 33342 and fixing them with paraformaldehyde. Images captured under the inverted fluorescence microscope are computationally processed with the specially designed ImageJ macro. Source:&nbsp;<strong>An effective &ldquo;three-in-one&rdquo; screening assay for testing drug and nanoparticle toxicity in human endothelial cells</strong> by Marcela Filipova et al., <em>PLOS</em>, Oct. 2018.
<strong>Time-response toxicity of different NPs towards HUVECs as measured by CDS assay.&nbsp;</strong>The HUVECs in the 96-well plate were treated with 100 &mu;g/ml of SPION, SiNP and CNTCOOH NPs for 0&ndash;24 h. The cell viability was measured by WST-8 assay (A), the cell necrosis was determined by LDH assay (B), and the number of intact cell nuclei (C) and number of apoptotic bodies (D) were counted by ImageJ software after the cells were stained with Hoechst 33342. Each treatment was performed in 6-plicate and the results (n = 3) are expressed as the means &plusmn; SEM as tested by one-way ANOVA followed by Dunnett&rsquo;s test. ***P&lt;0.001, **P&lt;0.01, and *P&lt;0.05, versus the time point 0 hours.&nbsp;Source:&nbsp;<strong>An effective &ldquo;three-in-one&rdquo; screening assay for testing drug and nanoparticle toxicity in human endothelial cells</strong>&nbsp;by Marcela Filipova et al.,&nbsp;<em>PLOS</em>, Oct. 2018.
<strong>Evaluation of CDS assay performance.</strong><br />The HUVECs in the 96-well plate were treated with different concentrations of camptothecin, staurosporine or H<sub>2</sub>O<sub>2</sub> for 24 hours (A&ndash;D), or with 5 &mu;M camptothecin, 100 nM staurosporine or 2 mM H<sub>2</sub>O<sub>2</sub> for 0, 3, 6, 12 and 24 hours (E&ndash;H). Cell viability was measured by WST-8 assay (A, E), and cell necrosis was evaluated by LDH assay (B, F). A count of the intact cell nuclei (C, G) and apoptotic bodies (D, H) was evaluated by ImageJ software after the cells were stained with Hoechst 33342. The WST-8 data and number of cell nuclei and apoptotic bodies were processed with 3h and 3.5h time difference, respectively. The data represent three independent experiments performed in 6-plicates. The bar graphs show the means &plusmn; SEM. Repeated measures were statistically tested by one-way ANOVA followed by Dunnett&rsquo;s post-test. ***P&lt;0.001, **P&lt;0.01, and *P&lt;0.05, versus the negative control.&nbsp;Source:&nbsp;<strong>An effective &ldquo;three-in-one&rdquo; screening assay for testing drug and nanoparticle toxicity in human endothelial cells</strong>&nbsp;by Marcela Filipova et al.,&nbsp;<em>PLOS</em>, Oct. 2018.

Documents

pdfCertificate of origin
pdfSafety data sheet
pdfProduct protocol
pdfCertificate of analysis
Cell Cycle Assays
Nucleus
DNA Methylation
Fluorescence Activated Cell Sorting (FACS)
Hoechst DNA Stains for Live and Fixed Cells
Immunohistochemistry (IHC)
Nucleic Acid Building Blocks

Alternative formats

Hoechst 33342 *20 mM solution in water*