Hoechst 33342 is a popular cell-permeant, blue fluorescent nuclear stain. It is used to visualize the nuclei of living or fixed cells and tissues and is often used to distinguish condensed, pyknotic nuclei in apoptotic cells. Hoechst 33342 emits blue fluorescence when bound to double stranded DNA. It is slightly more membrane permeant than the Hoechst 33258 analog. Hoechst 33342 may be used to identify healthy or apoptotic nuclear morphology and for cell cycle studies.
Hoechst 33342 Fluorescent Nucleic Acid Stain
Hoechst 33342 nucleic acid stain is a popular cell-permeant nuclear counterstain that emits blue fluorescence when bound to dsDNA. This dye is often used to distinguish condensed pyknotic nuclei in apoptotic cells and for cell cycle studies.
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1. Add Hoechst 33342 to the cell sample media at 0.5% v/v. For example, add 1.5 µL Hoechst to 300 µL of cells.
2. Incubate 10-20 minutes at room temperature.
3. Visualize with a fluorescence microscope by using a UV excitation and blue emission filter. The blue Hoechst stain fluoresces at 461 nm.
4. Alternatively, cells may be analyzed with a flow cytometer using a UV excitation source.
5. When bound to dsDNA, the maximum absorption is 350 nm and the maximum emission is 461.
• Supplied at: 200 µg/mL
• Excitation/Emission: 350 nm / 480 nm
• pH: 4.0 + 1.0
• Method of Analysis: Flow cytometer, Fluorescence Microscope
• Storage: 2-8°C
• Shipping: Ships overnight (domestic), International Priority Shipping
Czimmerer Z, Daniel B, Horvath A, Rückerl D, Nagy G, Kiss M, Peloquin M, Budai MM, Cuaranta-Monroy I, Simandi Z, Steiner L, Nagy B, Poliska S, Banko C, Bacso Z, Schulman IG, Sauer S, Deleuze JF, Allen JE, Benko S, Nagy L. The Transcription Factor STAT6 Mediates Direct Repression of Inflammatory Enhancers and Limits Activation of Alternatively Polarized Macrophages. Immunity. 2018. Jan 16;48(1):75-90.e6. doi: 10.1016/j.immuni.2017.12.010. Full text
“Sub-vital staining was performed in culture medium at room temperature for 20 minutes by Hoechst 34580 (10 microg/ml), propidium iodine (10 microg/ml), Alexa 488 tagged Annexin V (1 microg/ml) and caspase-1 specific FLICA® 660 (FLICA® 660 far-red fluorescence Caspase-1 Assay Kit was used according to the description of manufacturer; ImmunoChemistry Technologies, LLC).”