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1mg |
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5mg |
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10mg |
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ln Vitro |
Preparation of Hoechst working solution 1.1 Preparation of stock solution Use DMSO to prepare 1 mg/mL Hoechst stock solution. Note: Hoechst storage solution is recommended to be stored at -4℃ or -20℃ in the dark after filling. 1.2 Replacement of working solution: Inject excellent serum-free cell culture medium or PBS into the storage solution before use. The ultimate concentration is 10 μg/mL Hoechst working solution. Note: Please adjust the concentration of Hoechst working fluid according to the real condition, and make it suitable for usage. 2. Cell staining (suspended cells) 2.1 Collect cells by centrifugation, add PBS and wash twice, 5 minutes each time. When the cell density reaches 1×106/mL 2.2, add 1 mL Hoechst working solution for 3-10 minutes. 2.3 Centrifuge at 400 g for 3-4 minutes and discard. 2.4 Add PBS to wash the cells twice, 5 minutes each time. 2.5 Resuspend the cells in 1 mL of serum-free or PBS, and observe using a fluorescence microscope or flow cytometer. 3. Cell staining (adhesion) 3.1 Culture adherent cells on sterile coverslips. 3.2 Remove the coverslip from the culture medium and aspirate surplus cells. 3.3 Add 100 μL dye working solution, shake gently to completely cover the cells, leaving 3 3.4 Aspirate the dye working solution, wash 2-3 times with culture media, 5 minutes each time, and observe using a fluorescence microscope or flow cytometer. Notes 1. Please adjust the concentration of Hoechst working fluid according to the real conditions. -10 minutes. 2. This product is solely for scientific research by professionals and may not be used for clinical diagnosis or treatment, and may not be used in food or medicine. 3. For your safety and health, please use a lab coat and gloves when operating.
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ln Vivo |
Hoechst dye can bind to DNA in both fixed and live cells and is cell permeable [1].
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References |
[1]. a b c "Hoechst Stains". Invitrogren (Molecular Probes).
[2]. Latt SA, Stetten G, Juergens LA, Recent developments in the detection of deoxyribonucleic acid synthesis by 33258 Hoechst fluorescence. The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society 23 (7): 493-505. [3]. Portugal J, Waring MJ. Assignment of DNA binding sites for 4',6-diamidine-2-phenylindole and bisbenzimide (Hoechst 33258). A comparative footprinting study. Biochimica et Biophysica Acta 949 (2): 158-68. |
Molecular Formula |
C29H30N6O3
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Molecular Weight |
510.5869
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CAS # |
258843-62-8
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Related CAS # |
Hoechst 33258;23491-44-3;Hoechst 33258 analog 2;23491-54-5;Hoechst 33258 analog 3;23554-98-5;Hoechst 33258 analog 5;23491-55-6;Hoechst 33258 analog 6;129244-66-2
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Appearance |
Typically exists as solids (or liquids in special cases) at room temperature
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SMILES |
O(C([H])([H])C([H])([H])C([H])([H])C(=O)O[H])C1=C([H])C([H])=C([H])C(=C1[H])C1=NC2C([H])=C([H])C(=C([H])C=2N1[H])C1=NC2C([H])=C([H])C(=C([H])C=2N1[H])N1C([H])([H])C([H])([H])N(C([H])([H])[H])C([H])([H])C1([H])[H]
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HS Tariff Code |
2934.99.9001
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Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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Solubility (In Vitro) |
DMSO : ≥ 51 mg/mL (~99.88 mM)
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Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80: Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 :Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400  (Please use freshly prepared in vivo formulations for optimal results.) |
Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
1 mM | 1.9585 mL | 9.7926 mL | 19.5852 mL | |
5 mM | 0.3917 mL | 1.9585 mL | 3.9170 mL | |
10 mM | 0.1959 mL | 0.9793 mL | 1.9585 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration: mg/mL;
Method for preparing DMSO stock solution: mg drug pre-dissolved in μL DMSO (stock solution 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 stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O,mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.