Hoechst 33342: A Comprehensive Guide
Overview
Hoechst 33342 (bisbenzimide, HOE 33342) is a cell-permeant blue fluorescent dye widely used in biological research. It binds strongly to AT-rich regions in the minor groove of double-stranded DNA, enhancing its fluorescence. Known for its high cell permeability and lower toxicity compared to other dyes like DAPI, it is ideal for staining chromosomes and nuclei in live and fixed cells.
Spectral Properties
- Excitation Maximum: 351 nm
- Emission Maximum: 461 nm
- Stokes Shift: Large, ideal for multicolor labeling
- Unbound Dye Fluorescence: 510–540 nm (green)
Key Applications
- Fluorescence Microscopy: Stains chromosomes and nuclei for detailed imaging.
- Flow Cytometry: Differentiates cell populations, including apoptotic cells with condensed pycnotic nuclei.
- Cell Sorting: Identifies and separates live and dead cells in mixed samples.
Unique Features
- Cell Permeability: Hoechst 33342 is 10 times more permeable than Hoechst 33258 due to its ethyl group, allowing efficient staining of live cells.
- Compatibility with BrdU: The dye’s fluorescence is quenched by bromodeoxyuridine (BrdU), a tool for studying cell-cycle progression as BrdU incorporation disrupts optimal binding sites.
Concentrations and Usage
Commonly used at 0.1–10 μg/mL, Hoechst 33342 is effective for both bacteria and eukaryotic cells. Its application requires careful washing to avoid unbound dye interference, which can fluoresce green under certain conditions.
Product Availability
Hoechst 33342 is available in various forms, including powder (1H010), a 10 mg/mL concentrated solution (2G010), and an upcoming ready-to-use aqueous solution.
Conclusion
Hoechst 33342 is an essential tool in cell biology, offering high cell permeability, lower toxicity, and versatility in multicolor assays. Its unique properties make it valuable for research in cell cycle, apoptosis, and DNA visualization.
Explore our product options to find the right concentration and format for your research needs.
AxisPharm offers 5000+ PEG Linkers with high purity. Different kinds of PEG Reagents may be available by custom synthesis.
