Hoechst 33342

Tocris Bioscience | Catalog # 5117

Blue-fluorescent dye for DNA staining. Cell permeable. Used for: nuclear counterstain, apoptosis analysis, fixed and live-cell staining. Application: flow cytometry, confocal microscopy
Tocris Bioscience

Key Product Details

Description

Blue-fluorescent dye for DNA staining. Cell permeable. Used for: nuclear counterstain, apoptosis analysis, fixed and live-cell staining. Application: flow cytometry, confocal microscopy

Wavelength

Excitation = 350 nm, Emission = 461 nm
View all Fluorescent DNA Stains »

Product Description

Key information: Hoechst 33342 is a blue-fluorescent dye for DNA staining. Cell permeable. Suitable for fixed and live-cell staining. Commonly used as a counterstain in fluorescence microscopy.

Used for: nuclear counterstain, DNA visualization, cell cycle studies, apoptosis analysis.

Application: flow cytometry, confocal microscopy.

Properties and Photophysical Data: Hoechst 33342 binds to the AT-rich regions of the minor grove in DNA which renders it specific for nuclear chromatin. Excitation and emission maxima (λ) are 350 nm and 461 nm, respectively.

Scientific Data Images for Hoechst 33342

Fluorescent dye for staining DNA

Hoechst 33342 | CAS No. 875756-97-1

Fluorescent dye for staining DNA

Optical Data

Hoechst 33342 Dye Spectra
Emission Color Blue
λabs 350 nm
λem 461 nm
Cell Permeable Yes
Application Flow Cytometry, Confocal Microscopy

Spectra Viewer

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Use our spectra viewer to interactively plan your experiments, assessing multiplexing options. View the excitation and emission spectra for our fluorescent dye range and other commonly used dyes.

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Product Specifications for Hoechst 33342

Molecular Weight

561.93

Formula

C27H28N6O.3HCl

Storage

Store at -20°C

Purity

≥98% (HPLC)

Chemical Name

2'-(4-Ethoxyphenyl)-5-(4-methyl-1-piperazinyl)-2,5'-bi-1H-benzimidazole trihydrochloride

CAS Number

875756-97-1

PubChem ID

16760503

InChI Key

JABNPSKWVNCGMX-UHFFFAOYSA-N

SMILES

CN(CC2)CCN2C1=CC(N=C(C4=CC(NC(C6=CC=C(OCC)C=C6)=N5)=C5C=C4)N3)=C3C=C1.Cl.Cl.Cl

The technical data provided above is for guidance only. For batch specific data refer to the Certificate of Analysis.

Solubility

Solvent Max Conc. mg/mL Max Conc. mM
Solubility
water 28.1 50
DMSO 28.1 50

Preparing Stock Solutions for Hoechst 33342

The following data is based on the product molecular weight 561.93.

Batch specific molecular weights may vary from batch to batch due to the degree of hydration, which all affect the solvent volumes required to prepare stock solutions.

Select a batch to recalculate based on the batch molecular weight:

Concentration / Solvent Volume / Mass 1 mg 5 mg 10 mg
0.5 mM 3.56 mL 17.80 mL 35.59 mL
2.5 mM 0.71 mL 3.56 mL 7.12 mL
5 mM 0.36 mL 1.78 mL 3.56 mL
25 mM 0.07 mL 0.36 mL 0.71 mL

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Background References

References are publications that support the biological activity of the product. See our Citations tab to view 90 publications citing the usage of this product.

Product Documents for Hoechst 33342

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Product Specific Notices for Hoechst 33342

For research use only

Customer Reviews for Hoechst 33342 (7)

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Showing  1 - 5 of 7 reviews Showing All
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  • test
    Name: Chi Ngo
    Species: Mouse
    Assay Type: In Vivo
    Cell Line/Tissue: SH-SY5Y
    Verified Customer | Posted 09/06/2025
    Dilution: 1:1000. Hoechst 33342 was co-incubated with 2nd antibodies for 1h at RT,
    Hoechst 33342 was used to label nuclei of cells in ICC.
    Hoechst 33342 5117
  • HCT116 spheroids
    Name: Gabriele Strusi
    Species: Human
    Assay Type: In Vitro
    Verified Customer | Posted 12/12/2024
    HCT116 spheroids stained with 7 um Hoechst 33342
    Hoechst 33342 5117
  • MCF7 spheroids
    Name: Caterina Suelzu
    Species: Human
    Assay Type: In Vitro
    Cell Line/Tissue: MCF7
    Verified Customer | Posted 12/12/2024
    MCF7 spheroids stained with 7 uM Hoechst 33342
    Hoechst 33342 5117
  • Hoechst for nuclear staining
    Name: Anonymous
    Species: Human
    Assay Type: In Vitro
    Cell Line/Tissue: Macrophages
    Verified Customer | Posted 10/21/2021
    Hoechst was used for nuclear staining. Add stain at 1 ug/mL and incubate cells at room temperature for 10 minutes.
    Hoechst 33342 5117
  • staining nucleus of live cells
    Name: Anonymous
    Species: Human
    Assay Type: In Vitro
    Cell Line/Tissue: Chondrocytes
    Verified Customer | Posted 09/10/2020
    I used this dye to stain the nucleus of live cells. After making the stock solution, diluted the dye 1:1000 ratio in the culture media/PBS and added to cells and incubated for 5 mins.
    Hoechst 33342 5117
  • Staining nuclei
    Name: Anonymous
    Species: Human
    Assay Type: In Vitro
    Cell Line/Tissue: PC3
    Verified Customer | Posted 10/27/2019
    Add Hoechst dyes at 1 ug/mL.Incubate cells at room temperature for 10 minutes.
    Hoechst 33342 5117
  • Nuclear labeling
    Name: Anonymous
    Species: Human
    Assay Type: In Vitro
    Cell Line/Tissue: HEK 293
    Verified Customer | Posted 11/01/2017
    Works well at 1:2500 dilution incubated for 20 minutes at room temperature just before mounting the coverslips with the mounting media
    Immunostaining of HEK 293 cells
    Hoechst 33342 5117

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