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Hoechst 33342 *20 mM solution in water*

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 20 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)
Nanoengineered Neutrophil as 19F-MRI Tracer for Alert Diagnosis and Severity A of Acute Lung Injury
Authors: Li, Sha and Zhang, Lei and Xu, Qiuyi and Sui, Meiju and Xiao, Long and Chen, Daiqin and Jiang, Zhong-Xing and Zhou, Xin and Chen, Shizhen
Journal: Advanced Materials (2024): 2401513
The dynamin inhibitor, dynasore, prevents zoledronate-induced viability loss in human gingival fibroblasts by partially blocking zoledronate uptake and inhibiting endosomal acidification
Authors: Kirby, Jacob and Standfest, Makayla and Binkley, Jessica and Barnes, Charles and Brown, Evan and Cairncross, Tyler and Cartwright, Alex and Dadisman, Danielle and Mowat, Colten and Wilmot, Daniel and others,
Journal: Journal of Applied Oral Science (2024): e20240224
Mechanosensitive membrane domains regulate calcium entry in arterial endothelial cells to protect against inflammation
Authors: Hong, Soon-Gook and Ashby, Julianne W and Kennelly, John P and Wu, Meigan and Steel, Michelle and Chattopadhyay, Eesha and Foreman, Rob and Tontonoz, Peter and Tarling, Elizabeth J and Turowski, Patric and others,
Journal: The Journal of Clinical Investigation (2024)
Neuronal Glycogen Breakdown Mitigates Tauopathy via Pentose Phosphate Pathway-Mediated Oxidative Stress Reduction
Authors: Kapahi, Pankaj and Bar, Sudipta and Wilson, Kenneth and Hilsabeck, Tyler and Alderfer, Sydney and Dammer, Eric and Burton, Jordan and Shah, Samah and Holtz, Anja and Carrera, Enrique and others,
Journal: (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
Proton motive force-dependent Hoechst 33342 transport by the ABC transporter LmrA of Lactococcus lactis
Authors: van den Berg van Saparoea HB, Lubelski J, van Merkerk R, Mazurkiewicz PS, Driessen AJ.
Journal: Biochemistry (2005): 16931
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
Resistance mechanism development to the topoisomerase-I inhibitor Hoechst 33342 by Leishmania donovani
Authors: Marquis JF, Hardy I, Olivier M.
Journal: Parasitology (2005): 197
Page updated on November 21, 2024

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Catalog Number17535
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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™ (Cat No. 23002) working solution for 20 minutes in a 37°C, 5% CO<sub>2</sub> incubator and then washed 3 times. Nuclei were labeled with Hoechst 33342 (Cat No. 17530). Lysosomes were labeled with LysoBrite™ 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™ (Cat No. 23002) working solution for 20 minutes in a 37°C, 5% CO<sub>2</sub> incubator and then washed 3 times. Nuclei were labeled with Hoechst 33342 (Cat No. 17530). Lysosomes were labeled with LysoBrite™ 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™ (Cat No. 23002) working solution for 20 minutes in a 37°C, 5% CO<sub>2</sub> incubator and then washed 3 times. Nuclei were labeled with Hoechst 33342 (Cat No. 17530). Lysosomes were labeled with LysoBrite™ 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.
Gallery Image 3