Catalog | Size | Price | Quantity |
---|---|---|---|
17530 | 100 mg | Price | |
17533 | 1 g | Price |
Hoechst 33342 is a cell membrane-permeant, fluorescent DNA stain acclaimed for its strong binding affinity to A–T-rich regions in the minor groove of double-stranded DNA. This dye surpasses many nuclear stains due to its additional ethyl substituent, which boosts membrane permeability—thus seamlessly crossing the plasma membrane of live cells. As a result, Hoechst 33342 is widely utilized in diverse fields such as cell biology, immunology, neuroscience, and high-throughput drug discovery.
At AAT Bioquest, we deliver easy-to-use Hoechst 33342 solutions (equivalent to formulations from Sigma or Invitrogen). Built on decades of fluorescence chemistry expertise, our brand guarantees:
Chemical Family & Basic Properties
Note on Membrane Permeability: Structurally, Hoechst 33342 has an extra ethyl group for high membrane permeability (Kirby et al., 2024). Once bound to DNA, it emits intense blue fluorescence—ideal for a wide range of imaging and flow cytometry applications.
Excitation & Emission Characteristics
Its spectral overlap with DAPI allows reuse of existing filter sets. Flow cytometers with a 355 nm or 405 nm laser readily detect Hoechst 33342.
Working Concentration & Dosing Calculations
Given its 561.93 g/mol molecular weight, a 20 mM stock translates to ~11.2 mg/mL. Accurate calculations ensure reproducible staining intensities across experiments.
Key Advantages
Hoechst 33342 vs. DAPI
Hoechst 33342 is a non-intercalating dye that binds in the DNA minor groove—particularly in A–T–rich regions. This selective binding confers:
Why Fluorescence Increases Upon DNA Binding
In aqueous solution, free Hoechst 33342 exhibits a relatively low fluorescence quantum yield. However, once the dye inserts into the narrow A–T–rich minor groove:
Together, these factors can boost Hoechst 33342 fluorescence by up to 20–30×, making it exceptionally bright once bound to DNA (Lakowicz, 2006).
Additional Utility and Specificity
Researchers leverage these properties for a range of applications—from routine DNA quantification to probing DNA curvature and exploring conformational changes in complex assays.
Live-Cell and Fixed-Cell Nuclear Staining
Cell Cycle Analysis
Apoptosis and Cell Death Detection
High-Content Screening (HCS) and Automated Imaging
Side Population (SP) and Stem Cell Analysis
Additional Routine Applications
Recent Literature Highlights
Automated cell imaging [using Hoechst 33342] …include the ability to effectively compare adherent and suspension cell lines, reduced time to perform the assay, and environmental control allowing for long-term imaging studies. — Featherston et al., 2024
Experimental “Pearls”
Contrary to the prevailing assumption, Hoechst 33342 can be used in real-time imaging protocols for multiple days at sub-toxic concentrations, greatly expanding live-cell assay capacities. — Fuchs et al., 2023
Specialized & Emerging Applications
By leveraging these detailed insights on Hoechst 33342—from concentration handling to advanced imaging approaches—researchers can optimize assays across a spectrum of cell biology, microbiology, and therapeutic studies.
Molecular weight | 561.93 |
Solvent | Water |
Excitation (nm) | 352 |
Emission (nm) | 454 |
Certificate of Origin | Download PDF |
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 |
CAS | 875756-97-1 |
Fluorescence microscope | |
Excitation | 350 nm |
Emission | 461 nm |
Recommended plate | Black wall/clear bottom |
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