meta-iodoHoechst 33258 |
Catalog No.GC36586
One-Click Copy Product Info
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The nucleic acid stain meta-iodoHoechst 33258 is often used as a cell-permeable nuclear counterstain, which emits blue fluorescence after binding to dsDNA.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 158013-42-4
Sample solution is provided at 25 µL, 10mM.
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Hoechst stains are part of a family of blue fluorescent dyes used to stain DNA.IC50 Value:Target: These Bis-benzimides were originally developed by Hoechst AG, which numbered all their compounds so that the dye Hoechst 33342 is the 33342nd compound made by the company. There are three related Hoechst stains: Hoechst 33258, Hoechst 33342, and Hoechst 34580. The dyes Hoechst 33258 and Hoechst 33342 are the ones most commonly used and they have similarexcitation/emission spectra. Both dyes are excited by ultraviolet light at around 350 nm, and both emit blue/cyan fluorescent light around anemission maximum at 461 nm. Unbound dye has its maximum fluorescence emission in the 510-540 nm range. Hoechst dyes are soluble in water and in organic solvents such as dimethyl formamide or dimethyl sulfoxide. Concentrations can be achieved of up to 10 mg/mL. Aqueous solutions are stable at 2-6 °C for at least six months when protected from light. For long-term storage the solutions are instead frozen at ≤-20 °C.The dyes bind to the minor groove of double-stranded DNA with a preference for sequences rich in adenine andthymine. Although the dyes can bind to all nucleic acids, AT-rich double-stranded DNA strands enhance fluorescence considerably.Hoechst dyes are cell-permeable and can bind to DNA in live or fixed cells. Therefore, these stains are often called supravital, which means that cells survive a treatment with these compounds. Cells that express specific ATP-binding cassette transporter proteins can also actively transport these stains out of their cytoplasm.in vitro: N/A in vivo: N/AClinical trial: N/A
[1]. 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. [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]. a b c "Hoechst Stains". Invitrogren (Molecular Probes).
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 solid and prepare a 10 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 10 μg/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 5-10 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 100uL 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 5-15 minutes.
(4) Aspirate away the dye working solution, wash the coverslip 2 to 3 times with culture solution, and observe with a fluorescence microscope.
Precautions:
① For staining of fixed cells or tissue samples, the fixative needs to be removed by rinsing after fixation;
② Hoechst 33258 staining is usually performed after other staining. If no other staining is required, Hoechst 33258 staining is performed directly;
③ To slow down fluorescence quenching, it is recommended to use anti-fluorescence quenching mounting agent;
④ Fluorescent dyes all have quenching problems, so please try to avoid light;
⑤ Hoechst 33258 is irritating to the human body. For your safety and health, please wear a lab coat and disposable gloves when operating.
References:
[1]. K D Harshman, P B Dervan. Molecular recognition of B-DNA by Hoechst 33258. 1985 Jul 11;13(13):4825-35. doi: 10.1093/nar/13.13.4825.
| Cas No. | 158013-42-4 | SDF | |
| Canonical SMILES | CN1CCN(C2=CC=C3N=C(C4=CC=C5N=C(C6=CC=CC(I)=C6)NC5=C4)NC3=C2)CC1 | ||
| Formula | C25H23IN6 | M.Wt | 534.39 |
| Solubility | Soluble in DMSO | Storage | 4°C, protect from light |
| 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 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 1.8713 mL | 9.3565 mL | 18.7129 mL |
| 5 mM | 374.3 μL | 1.8713 mL | 3.7426 mL |
| 10 mM | 187.1 μL | 935.6 μL | 1.8713 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
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 saline, 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: >98.00% Appearance: A solid
- COA (Certificate of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
Average Rating: 5 (Based on Reviews and 2 reference(s) in Google Scholar.)