Hoechst 33258 analog 5 |
Catalog No.GC36242
One-Click Copy Product Info
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Hoechst 33258 analog 5 is a DNA minor groove-binding fluorescent dye belonging to the Hoechst series of labeling dyes. Hoechst 33258 analog 5 binds to A/T-rich regions of DNA and emits strong blue fluorescence (excitation/emission wavelengths = 350-365nm/460-490nm).
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 23491-55-6
Sample solution is provided at 25 µL, 10mM.
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Hoechst 33258 analog 5 is a DNA minor groove-binding fluorescent dye belonging to the Hoechst series of labeling dyes. Hoechst 33258 analog 5 binds to A/T-rich regions of DNA and emits strong blue fluorescence (excitation/emission wavelengths = 350-365nm/460-490nm). Hoechst 33258 analog 5 can be used in research related to cell cycle analysis, chromatin structure studies, high-throughput screening, and apoptosis detection[1-4].
References:
[1] Xing C, Liu J, Chen F, et al. Diphenyl-1-pyrenylphosphine: photo-triggered AIE/ACQ transition with remarkable third-order nonlinear optical signal change. Chem Commun (Camb). 2020 Apr 14;56(30):4220-4223.
[2] Santas J, Guardiola F, Rafecas M, et al. Determination of total plasma hydroperoxides using a diphenyl-1-pyrenylphosphine fluorescent probe. Anal Biochem. 2013 Mar 1;434(1):172-7.
[3] Latt SA, Stetten G, Juergens LA, et al. Recent developments in the detection of deoxyribonucleic acid synthesis by 33258 Hoechst fluorescence. J Histochem Cytochem. 1975 Jul;23(7):493-505.
[4] Singh MP, Joseph T, Kumar S, et al. Synthesis and sequence-specific DNA binding of a topoisomerase inhibitory analog of Hoechst 33258 designed for altered base and sequence recognition. Chem Res Toxicol. 1992 Sep-Oct;5(5):597-607.
Hoechst 33258 analog 5 Staining Protocol[1]
This protocol is adapted from research data and provided for reference only. Adjustments may be necessary based on specific experimental requirements.
1. Preparation of Staining Solution
(1) Prepare Stock Solution: Dissolve Hoechst 33258 analog 5 in DMSO or deionized water to prepare a stock solution with a concentration of 1-5mM. Note: Aliquot unused stock solution and store protected from light at -20°C or -80°C. Avoid repeated freeze-thaw cycles.
(2) Prepare Working Solution: Dilute the stock solution with a suitable buffer (e.g., PBS, serum-free medium, or HBSS) to prepare a working solution with a concentration of 1-10µM (commonly used range is 1-5µM). Note: Adjust the working solution concentration according to the actual situation. Prepare fresh before use.
2. Staining of Suspension Cells
(1) Suspension Cells: Centrifuge at 4°C, 1000g for 3-5 minutes. Discard the supernatant and wash twice with PBS.
(2) Adherent Cells: Wash twice with PBS, then digest cells with trypsin. After digestion is complete, centrifuge at 1000g for 3-5 minutes.
(3) Resuspend the cell pellet in 1 mL of Hoechst 33258 analog 5 working solution. Incubate protected from light at 37°C for 15-30 minutes. The optimal incubation time varies for different cell types; please optimize based on specific experimental requirements.
(4) After incubation, centrifuge at 1000g for 5 minutes. Discard the supernatant and wash the cell pellet with PBS 2-3 times.
(5) Resuspend cells in pre-warmed serum-free cell culture medium or PBS. Observe using fluorescence microscopy or flow cytometry.
3. Staining of Adherent Cells
(1) Culture adherent cells on sterile coverslips.
(2) Remove the coverslip from the culture medium, aspirate excess medium, and place the coverslip in a humid environment.
(3) Add an appropriate amount of Hoechst 33258 analog 5 working solution (typically 100-500µL, ensuring coverage of all cells). Gently rock to distribute the dye evenly.
(4) Incubate protected from light at 37°C for 15-30 minutes. The optimal incubation time varies for different cell types; please optimize based on specific experimental requirements.
(5) After incubation, aspirate the dye working solution. Wash the coverslip 2-3 times with pre-warmed culture medium.
4. Microscopy Detection: The excitation/emission wavelengths of Hoechst 33258 analog 5 are approximately 350-365nm/460-490nm (blue fluorescence). Use a DAPI filter set for observation.
Precautions:
(1) It is recommended to set up positive and negative controls (e.g., cells without dye).
(2) All fluorescent dyes are subject to photobleaching. Please protect samples from light as much as possible to slow down fluorescence quenching.
(3) Hoechst 33258 analog 5 dye is cytotoxic. Prolonged incubation may affect cell viability. Hoechst 33258 analog 5 is recommended to perform detection as soon as possible after staining.
(4) For your safety and health, please wear a lab coat and disposable gloves during operation.
References:
[1] Reiser O. Shining Light on Copper: Unique Opportunities for Visible-Light-Catalyzed Atom Transfer Radical Addition Reactions and Related Processes. Acc Chem Res. 2016 Sep 20;49(9):1990-6.
| Cas No. | 23491-55-6 | SDF | |
| Canonical SMILES | CN1CCN(C2=CC=C3N=C(C4=CC=C5N=C(C6=CC=C7C=CC=CC7=C6)NC5=C4)NC3=C2)CC1 | ||
| Formula | C29H26N6 | M.Wt | 458.56 |
| Solubility | DMSO : 33.33 mg/mL (72.68 mM; Need ultrasonic) | 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 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 2.1807 mL | 10.9037 mL | 21.8074 mL |
| 5 mM | 436.1 μL | 2.1807 mL | 4.3615 mL |
| 10 mM | 218.1 μL | 1.0904 mL | 2.1807 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 4 reference(s) in Google Scholar.)