Hoechst 34580 tetrahydrochloride (Synonyms: HOE 34580 tetrahydrochloride) |
| Catalog No.GC36247 |
El tetrahidrocloruro de Hoechst 34580 es un tinte fluorescente permeable a las células para teÑir ADN y nÚcleos.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 2310135-08-9
Sample solution is provided at 25 µL, 10mM.
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Hoechst 34580 tetrahydrochloride is a Hoechst series dye that is commonly used for nuclear labeling and staining of living cells. The maximum excitation/emission light is 392⁄490nm respectively. Hoechst dyes can be used to monitor cell viability by tracking changes in their emission spectra. As slightly groove-binding DNA stains with AT selectivity, Hoechst dyes are able to bind to all nucleic acids, but they show greater fluorescence enhancement for AT-rich double-stranded DNA strands compared to GC-rich strands[1]. This property has been used to identify Q bands in chromosomes, which are AT base pair-rich regions that fluoresce brightly when stained with quinacrine dye[2]. The fluorescence intensity of Hoechst dye increases as the pH of the solution increases.
Hoechst 34580 tetrahydrochloride is commonly used as a cell-permeable nuclear counterstain that emits blue fluorescence upon binding to dsDNA. Hoechst 34580 tetrahydrochloride is also used in various studies related to cell counting, cell cycle analysis and cell replication. It is particularly useful for identifying condensed nuclei in apoptotic cells and for cell cycle studies in combination with BrdU staining.
References:
[1].Karen E. Selph. Enumeration of marine microbial organisms by flow cytometry using near-UV excitation of Hoechst 34580-stained DNA.
[2]. Jonas Bucevičius,Gražvydas Lukinavičius , Rūta Gerasimaitė. The Use of Hoechst Dyes for DNA Staining and Beyond. Chemosensors 2018, 6(2), 18. Doi:org/10.3390/chemosensors6020018.
This plan only provides a guide, please modify it to meet your specific needs.
1. Prepare Hoechst staining solution
(1) Prepare Hoechst dye stock solution: Use DMSO to dissolve the solid powder and prepare a 1-5 mg/mL Hoechst dye stock solution.
Note: Hoechst stock solution is recommended to be aliquoted and stored in the dark at -4°C or -20°C to avoid repeated freezing and thawing.
(2) Working solution preparation: Use preheated serum-free medium or buffer (such as HBSS or PBS) to dilute the stock solution and prepare a Hoechst working solution with a concentration of 1-10μg/mL.
Note: Please adjust the concentration of Hoechst working fluid according to the actual situation and prepare it immediately.
2. Cell staining
2.1 Suspension cells (taking 6-well plate as an example)
(1) Centrifuge suspended cells at 1000g for 3-5 minutes. Discard the supernatant and wash twice with PBS for 5 minutes each time.
(2) Add 1 mL of Hoechst dye working solution and incubate at room temperature in the dark for 5-30 minutes.
(3) After the incubation, centrifuge at 1000g for 5 minutes, remove the supernatant, add PBS and wash 2-3 times, 5 minutes each time.
(4) Use serum-free cell culture medium or PBS to resuspend the cells and observe them through fluorescence microscopy or flow cytometry.
2.2 Adherent cells
(1) Culture adherent cells on sterile coverslips.
(2) Remove the coverslip from the culture medium, suck out the excess culture medium, and place the coverslip in a humid environment.
(3) Add 100μL of Hoechst dye working solution from one corner of the coverslip, shake gently to evenly cover all cells with the dye, and incubate at room temperature in the dark for 5-30 minutes.
(4) Aspirate away the dye working solution, wash the coverslip 2 to 3 times with culture solution, and observe with a fluorescence microscope.
3. Microscope detection: Hoechst 34580 can be excited by near-UV (375nm) or purple (405nm) laser, and the maximum excitation/emission light is 392⁄490nm respectively[1][2].
Precautions:
1) Fluorescent dyes all have quenching problems. Please avoid light as much as possible to slow down fluorescence quenching.
2) For your safety and health, please wear a lab coat and disposable gloves.
References:
[1].Karen E. Selph. Enumeration of marine microbial organisms by flow cytometry using near-UV excitation of Hoechst 34580-stained DNA.
[2]. Jonas Bucevičius,Gražvydas Lukinavičius , Rūta Gerasimaitė. The Use of Hoechst Dyes for DNA Staining and Beyond. Chemosensors 2018, 6(2), 18. Doi:org/10.3390/chemosensors6020018.
| Cas No. | 2310135-08-9 | SDF | |
| Sinónimos | HOE 34580 tetrahydrochloride | ||
| Canonical SMILES | CN1CCN(C2=CC=C3C(N=C(C4=CC=C5C(N=C(C6=CC=C(N(C)C)C=C6)N5)=C4)N3)=C2)CC1.[H]Cl.[H]Cl.[H]Cl.[H]Cl | ||
| Formula | C27H33Cl4N7 | M.Wt | 597.41 |
| Solubility | Water: 2.67 mg/mL (4.47 mM) | Storage | Store at -20°C |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
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| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
|
1 mg | 5 mg | 10 mg |
| 1 mM | 1.6739 mL | 8.3695 mL | 16.7389 mL |
| 5 mM | 0.3348 mL | 1.6739 mL | 3.3478 mL |
| 10 mM | 0.1674 mL | 0.8369 mL | 1.6739 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
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μL
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Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >99.50%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 5 reference(s) in Google Scholar.)
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