Hoechst 33342 *20 mM solution in water*
Ordering information
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Additional ordering information
Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
sales@aatbio.com | |
International | See distributors |
Bulk request | Inquire |
Custom size | Inquire |
Shipping | Standard overnight for United States, inquire for international |
Physical properties
Molecular weight | 561.93 |
Solvent | Water |
Spectral properties
Excitation (nm) | 352 |
Emission (nm) | 454 |
Storage, safety and handling
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 |
Alternative formats
Hoechst 33342 *Ultrapure Grade* |
Overview | ![]() ![]() |
See also: Cell Cycle Assays, Nucleus, Fluorescence Activated Cell Sorting (FACS), Hoechst DNA Stains for Live and Fixed Cells, Immunohistochemistry (IHC)
CAS 23491-52-3 | Molecular weight 561.93 | Excitation (nm) 352 | Emission (nm) 454 |
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.
Platform
Fluorescence microscope
Excitation | 350 nm |
Emission | 461 nm |
Recommended plate | Black wall/clear bottom |
Calculators
Common stock solution preparation
Table 1. Volume of Water needed to reconstitute specific mass of Hoechst 33342 *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.
0.1 mg | 0.5 mg | 1 mg | 5 mg | 10 mg | |
1 mM | 177.958 µL | 889.791 µL | 1.78 mL | 8.898 mL | 17.796 mL |
5 mM | 35.592 µL | 177.958 µL | 355.916 µL | 1.78 mL | 3.559 mL |
10 mM | 17.796 µL | 88.979 µL | 177.958 µL | 889.791 µL | 1.78 mL |
Molarity calculator
Enter any two values (mass, volume, concentration) to calculate the third.
Mass (Calculate) | Molecular weight | Volume (Calculate) | Concentration (Calculate) | Moles | ||||
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Product Family
Name | Excitation (nm) | Emission (nm) | Extinction coefficient (cm -1 M -1) | Quantum yield |
Hoechst 33258 *CAS 23491-45-4* | 352 | 454 | 460001 | 0.03401 |
Hoechst 33258 *20 mM solution in water* | 352 | 454 | 460001 | 0.03401 |
Hoechst 34580 *CAS 911004-45-0* | 371 | 438 | - | - |
Hoechst 34580 *20 mM solution in water* | 371 | 438 | - | - |
Images

Figure 1. 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% CO2 incubator and then washed 3 times. Nuclei were labeled with Hoechst 33342 (Cat No. 17530). Lysosomes were labeled with LysoBrite™ Orange (Cat No. 22657).

Figure 2. HeLa cells were seeded in 96-well microplates and incubated at 37 °C, 5% CO2 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.
Citations
View all 15 citations: Citation Explorer
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)
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)
Repair Kinetics of DSB-Foci Induced by Proton and $\alpha$-Particle Microbeams of Different Energies
Authors: Belchior, Ana and Canhoto, Jo{\~a}o F and Giesen, Ulrich and Langner, Frank and Rabus, Hans and Schulte, Reinhard
Journal: Life (2022): 2040
Authors: Belchior, Ana and Canhoto, Jo{\~a}o F and Giesen, Ulrich and Langner, Frank and Rabus, Hans and Schulte, Reinhard
Journal: Life (2022): 2040
Repair kinetics of DSB-foci induced by proton and helium ion microbeams of different energies
Authors: Belchior, Ana and Canhoto, Jo{\~a}o F and Giesen, Ulrich and Langner, Frank and Rabus, Hans and Schulte, Reinhard
Journal: arXiv preprint arXiv:2206.08981 (2022)
Authors: Belchior, Ana and Canhoto, Jo{\~a}o F and Giesen, Ulrich and Langner, Frank and Rabus, Hans and Schulte, Reinhard
Journal: arXiv preprint arXiv:2206.08981 (2022)
Aptamerized silica/gold nanocapsules for stimulated release of doxorubicin through remote two-photon excitation
Authors: Tew, Lih Shin and Lee, Tsung-Hsi and Lo, Leu-Wei and Khung, Yit Lung and Chen, Nai-Tzu
Journal: International Journal of Smart and Nano Materials (2022): 79--99
Authors: Tew, Lih Shin and Lee, Tsung-Hsi and Lo, Leu-Wei and Khung, Yit Lung and Chen, Nai-Tzu
Journal: International Journal of Smart and Nano Materials (2022): 79--99
NAMPT Inhibitor and P73 Activator Represses P53 R175H Mutated HNSCC Cell Proliferation in a Synergistic Manner
Authors: Cai, Bi-He and Bai, Zhi-Yu and Lien, Ching-Feng and Yu, Si-Jie and Lu, Rui-Yu and Wu, Ming-Han and Wu, Wei-Chen and Chen, Chia-Chi and Hsu, Yi-Chiang
Journal: Biomolecules (2022): 438
Authors: Cai, Bi-He and Bai, Zhi-Yu and Lien, Ching-Feng and Yu, Si-Jie and Lu, Rui-Yu and Wu, Ming-Han and Wu, Wei-Chen and Chen, Chia-Chi and Hsu, Yi-Chiang
Journal: Biomolecules (2022): 438
Studium bun{\v{e}}{\v{c}}n{\'e} toxicity vybran{\`y}ch nano{\v{c}}{\'a}stic v tk{\'a}{\v{n}}ov{\`y}ch kultur{\'a}ch.
Authors: Filipov{\'a}, Marcela
Journal: (2020)
Authors: Filipov{\'a}, Marcela
Journal: (2020)
Thoroughly remold the localization and signaling pathway of TLR22
Authors: Ji, Jianfei and Liao, Zhiwei and Rao, Youliang and Li, Wenqian and Yang, Chunrong and Yuan, Gailing and Feng, Hao and Xu, Zhen and Shao, Jianzhong and Su, Jianguo
Journal: Frontiers in immunology (2020): 3003
Authors: Ji, Jianfei and Liao, Zhiwei and Rao, Youliang and Li, Wenqian and Yang, Chunrong and Yuan, Gailing and Feng, Hao and Xu, Zhen and Shao, Jianzhong and Su, Jianguo
Journal: Frontiers in immunology (2020): 3003
Plant-derived alkaloid sinomenine potentiates glucocorticoid pharmacodynamics in mitogen-activated human peripheral blood mononuclear cells by regulating the translocation of glucocorticoid receptor
Authors: Xu, Wencheng and Wang, Xiaoqin and Tu, Yuanchao and Masaki, Hiroshi and Tanaka, Sachiko and Onda, Kenji and Sugiyama, Kentaro and Yamada, Haruki and Hirano, Toshihiko
Journal: Phytotherapy research (2019): 187--196
Authors: Xu, Wencheng and Wang, Xiaoqin and Tu, Yuanchao and Masaki, Hiroshi and Tanaka, Sachiko and Onda, Kenji and Sugiyama, Kentaro and Yamada, Haruki and Hirano, Toshihiko
Journal: Phytotherapy research (2019): 187--196
Tetrandrine and cepharanthine induce apoptosis through caspase cascade regulation, cell cycle arrest, MAPK activation and PI3K/Akt/mTOR signal modification in glucocorticoid resistant human leukemia Jurkat T cells
Authors: Xu, Wencheng and Wang, Xiaoqin and Tu, Yuanchao and Masaki, Hiroshi and Tanaka, Sachiko and Onda, Kenji and Sugiyama, Kentaro and Yamada, Haruki and Hirano, Toshihiko
Journal: Chemico-Biological Interactions (2019): 108726
Authors: Xu, Wencheng and Wang, Xiaoqin and Tu, Yuanchao and Masaki, Hiroshi and Tanaka, Sachiko and Onda, Kenji and Sugiyama, Kentaro and Yamada, Haruki and Hirano, Toshihiko
Journal: Chemico-Biological Interactions (2019): 108726
Helicobacter pylori cholesterol glucosylation modulates autophagy for increasing intracellular survival in macrophages
Authors: Lai, Chih-Ho and Huang, Ju-Chun and Cheng, Hsin-Hung and Wu, Meng-Chen and Huang, Mei-Zi and Hsu, Hui-Ying and Chen, Yu-An and Hsu, Chung-Yao and Pan, Yi-Jiun and Chu, Yen-Ting and others, undefined
Journal: Cellular microbiology (2018): e12947
Authors: Lai, Chih-Ho and Huang, Ju-Chun and Cheng, Hsin-Hung and Wu, Meng-Chen and Huang, Mei-Zi and Hsu, Hui-Ying and Chen, Yu-An and Hsu, Chung-Yao and Pan, Yi-Jiun and Chu, Yen-Ting and others, undefined
Journal: Cellular microbiology (2018): e12947
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
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
Authors: Zhang X, Kiechle FL.
Journal: Ann Clin Lab Sci (2006): 185
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
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
Authors: Marquis JF, Hardy I, Olivier M.
Journal: Parasitology (2005): 197
Acid-base and electronic structure-dependent properties of Hoechst 33342
Authors: Aleman C, Namba AM, Casanovas J.
Journal: J Biomol Struct Dyn (2005): 29
Authors: Aleman C, Namba AM, Casanovas J.
Journal: J Biomol Struct Dyn (2005): 29
Single UV excitation of Hoechst 33342 and propidium iodide for viability assessment of rhesus monkey spermatozoa using flow cytometry
Authors: Cai K, Yang J, Guan M, Ji W, Li Y, Rens W.
Journal: Arch Androl (2005): 371
Authors: Cai K, Yang J, Guan M, Ji W, Li Y, Rens W.
Journal: Arch Androl (2005): 371
Evidence for a qualitative hierarchy within the Hoechst-33342 'side population' (SP) of murine bone marrow cells
Authors: Robinson SN, Seina SM, Gohr JC, Kuszynski CA, Sharp JG.
Journal: Bone Marrow Transplant (2005): 807
Authors: Robinson SN, Seina SM, Gohr JC, Kuszynski CA, Sharp JG.
Journal: Bone Marrow Transplant (2005): 807
Flow cytometric characterization of viable meiotic and postmeiotic cells by Hoechst 33342 in mouse spermatogenesis
Authors: Bastos H, Lassalle B, Chicheportiche A, Riou L, Testart J, Allem and I, Fouchet P.
Journal: Cytometry A (2005): 40
Authors: Bastos H, Lassalle B, Chicheportiche A, Riou L, Testart J, Allem and I, Fouchet P.
Journal: Cytometry A (2005): 40
An in vitro study of Hoechst 33342 redistribution and its effects on cell viability
Authors: Mohorko N, Kregar-Velikonja N, Repovs G, Gorensek M, Bresjanac M.
Journal: Hum Exp Toxicol (2005): 573
Authors: Mohorko N, Kregar-Velikonja N, Repovs G, Gorensek M, Bresjanac M.
Journal: Hum Exp Toxicol (2005): 573
Role of topoisomerases in cytotoxicity induced by DNA ligand Hoechst-33342 and UV-C in a glioma cell line
Authors: Singh S, Dwarakanath BS, Lazar Mathew T.
Journal: Indian J Exp Biol (2005): 313
Authors: Singh S, Dwarakanath BS, Lazar Mathew T.
Journal: Indian J Exp Biol (2005): 313