Hoechst 33342 Staining Solution (1 mg/mL), IC-20015
Product Introduction
This product is a ready-to-use fluorescent nuclear stain containing Hoechst 33342 at a concentration of 1 mg/mL. Hoechst 33342 is a cell-permeable, blue fluorescent dye that binds specifically to the minor groove of double-stranded DNA, with a preference for adenine-thymine (A-T) rich regions. Upon binding to DNA, the dye exhibits a significant increase in fluorescence intensity, enabling sensitive detection and quantification of nuclear material in both live and fixed cells.
Hoechst 33342 is widely used for nuclear counterstaining in immunofluorescence microscopy, cell cycle analysis by flow cytometry, apoptosis detection, and chromatin condensation assessment. Its cell permeability allows for staining of live cells without the need for permeabilization, making it ideal for time-lapse imaging and live-cell tracking applications. The dye is compatible with a broad range of fluorescent probes including GFP, FITC, and rhodamine derivatives.
Product Features
1. Cell-Permeable: Stains nuclei in live cells without fixation or permeabilization, suitable for time-lapse and live-cell imaging.
2. High Affinity for DNA: Exhibits strong fluorescence enhancement upon binding to A-T rich regions of double-stranded DNA.
3. Ready-to-Use 1 mg/mL Solution: Supplied as a sterile-filtered stock solution for convenient dilution to working concentrations.
4. Versatile Applications: Compatible with flow cytometry, fluorescence microscopy, high-content screening, and apoptosis assays.
5. Stable Fluorescence: Provides bright, photostable blue nuclear staining for extended observation periods.
Specifications
Size: Available in 1 mL and 10¡Á1 mL
Concentration: 1 mg/mL in sterile deionized water
Storage and Stability
Storage Conditions: Store at -20¡ãC, protected from light. For frequent use, aliquot and store at 4¡ãC for up to 1 month.
Shelf Life: The product is stable for 12 months from the date of manufacture when stored as directed. Avoid repeated freeze-thaw cycles.
Excitation/Emission: 350 nm / 461 nm (Blue Fluorescence upon DNA binding)
Protocol (For Reference Only)
Important: Protect dye from light during handling and storage. Optimize working concentration for specific cell types and experimental conditions.
1. Live Cell Staining: Dilute Hoechst 33342 stock solution directly into complete culture medium to a final concentration of 1-10 µg/mL (1-10 µL per mL medium), incubate cells at 37¡ãC for 10-30 minutes, then proceed directly to imaging without washing.
2. Fixed Cell Staining: Prepare fixed and permeabilized cells according to standard immunofluorescence protocols, dilute Hoechst 33342 stock solution in PBS to a final concentration of 0.1-1 µg/mL, incubate at room temperature for 5-10 minutes, and wash briefly with PBS before mounting and imaging.
3. Flow Cytometry: Dilute Hoechst 33342 stock solution in cell suspension buffer to a final concentration of 1-5 µg/mL, incubate at 37¡ãC for 15-30 minutes, and analyze using UV laser excitation (355 nm) with a 450/50 nm bandpass filter.
4. Apoptosis and Cell Cycle Analysis: Use Hoechst 33342 in combination with propidium iodide (PI) for viability discrimination or as a standalone DNA content stain for cell cycle profiling.
5. Imaging: Visualize using a DAPI filter set with UV excitation and blue emission collection.
Precautions
1. Protect from light during handling and storage to prevent photobleaching and loss of fluorescence.
2. Optimize staining concentration for each cell type as dye uptake and efflux rates vary among different cell lines due to multidrug resistance transporter activity.
3. Avoid prolonged exposure to high dye concentrations in live cells as Hoechst 33342 may exhibit cytotoxicity and affect cell cycle progression at elevated doses.
4. For fixed cells, ensure complete removal of excess dye by washing to minimize background fluorescence.
5. For research use only. Not for use in diagnostic or therapeutic procedures.
FAQ (Simplified)
Q1: What is the difference between Hoechst 33342 and Hoechst 33258?
A1: Hoechst 33342 is more lipophilic and therefore more cell-permeable than Hoechst 33258, making it the preferred choice for live-cell staining applications. Hoechst 33258 is typically used for fixed cell staining.
Q2: Why is my staining intensity variable between different cell types?
A2: Hoechst 33342 is a substrate for multidrug resistance (MDR) efflux pumps. Cells expressing high levels of ABC transporters may actively pump out the dye, requiring higher dye concentrations or co-incubation with efflux inhibitors.
Q3: Can I use Hoechst 33342 with GFP-expressing cells?
A3: Yes. Hoechst 33342 excites in the UV range and emits blue fluorescence, which is spectrally separated from GFP (green) emission. This makes it an excellent nuclear counterstain for GFP co-localization studies.
Q4: How should I store the product after opening?
A4: For long-term storage, aliquot the stock solution into smaller volumes and store at -20¡ãC protected from light. Avoid repeated freeze-thaw cycles. A working aliquot may be stored at 4¡ãC for up to one month.
Disclaimer
1. For Research Use Only. Not for use in diagnostic or therapeutic procedures.
2. Due to the variable nature of biological research, optimization of conditions using a small-scale pilot experiment is strongly recommended before processing valuable samples.
3. This warranty is limited to the replacement of the product. The manufacturer assumes no liability for incidental or consequential damages, including loss of samples or data.
4. Wear appropriate protective clothing and gloves when handling this product.
Ordering Information
Catalog Number: IC-20015
Product Name: Hoechst 33342 Staining Solution (1 mg/mL)
Size: 1 mL / 10¡Á1 mL
Price:
1 mL: CNY ¥600.00 / USD $60.00 / EUR €72.00 / JPY ¥10800.00
10¡Á1 mL: CNY ¥4500.00 / USD $450.00 / EUR €540.00 / JPY ¥81000.00
2023 Version
