DAPI (hydrochloride) (Synonyms: 4',6Diamidino2phenylindole, FxCycle Violet) |
Catalog No.GC43378
One-Click Copy Product Info
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DAPI (hydrochloride), as a DNA-specific probe, can form a fluorescent complex by attaching in the minor grove of A-T rich sequences of DNA and nonfluorescent intercalative complexes with double-stranded nucleic acids.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 28718-90-3
Sample solution is provided at 25 µL, 10mM.
- Neuropharmacology 297 (2026):111051.PMID:42214456
- Neurotoxicology 88 (2022):134-143.PMID:34785253
- Am j transl res 14.10 (2022):7012.PMID:36398229
- J Virol (2025):e00352-25.PMID:40172227
- Ecotox Environ Safe 213 (2021):112040.PMID:33610943
- J Inorg Biochem 230 (2022):111750.PMID:35151098
- Evid Based Compl Alt (2022).
- Chin J Exp Ophthalmol (2023):127-133.
- bioRxiv (2026):2026-01.
DAPI (hydrochloride), as a DNA-specific probe, can form a fluorescent complex by attaching in the minor grove of A-T rich sequences of DNA and nonfluorescent intercalative complexes with double-stranded nucleic acids. DAPI, as a chromosome and nuclear stain, has maximum excitation ultraviolet (UV) light wavelength with 358nm and emission in the blue range with 461nm[1]. DAPI is usually used for flow cytometry, chromosome staining, DNA visualization and quantitation in histochemistry and biochemistry[2]. DAPI is a good substrate of hOCT1 with a Michaelis constant of 8.94µM[3]. 1.43µM DAPI can measure mitochondrial permeability transition and mitochondrial membrane depolarization by combining with Annexin V (FITC) and the potentiometric fluorescent dye, tetramethylrhodamine methyl ester (TMRM)[4].
References:
[1] Karg TJ, et al. Photoconversion of DAPI and Hoechst dyes to green and red-emitting forms after exposure to UV excitation. Chromosoma. 2018 Jun;127(2):235-245.
[2] Kapuscinski J. DAPI: a DNA-specific fluorescent probe. Biotech Histochem. 1995 Sep;70(5):220-33.
[3] Yasujima T, et al. Characterization of human OCT1-mediated transport of DAPI as a fluorescent probe substrate. J Pharm Sci. 2011 Sep;100(9):4006-12.
[4] Wallberg F, et al. Analysis of Apoptosis and Necroptosis by Fluorescence-Activated Cell Sorting. Cold Spring Harb Protoc. 2016 Apr 1;2016(4):pdb.prot087387.
This plan only provides a guide, please modify it to meet your specific needs.
1. Preparation of staining solution
(1) Prepare dye stock solution Use ddH2O to dissolve DAPI and prepare a stock solution with a concentration of 1-10mgml.
Notice
① Do not use buffer to dissolve the powder directly;
② The prepared stock solution needs to be filtered and sterilized using a 0.22μm filter membrane;
③ Unused stock solution should be aliquoted according to single dosage and stored in the dark at -20℃ or -80℃ to avoid repeated freezing and thawing.
(2) Prepare dye working solution Use ddH2O or PBS to dilute the stock solution and prepare a working solution with a concentration of 0.5-10μgml.
Note Please adjust the concentration of the working fluid according to the actual situation and prepare it now.
2. Cell suspension staining
(1) Suspension cells Centrifuge at 1000g for 3-5 minutes at 4℃, discard the supernatant, and wash twice with PBS for 5 minutes each time.
(2) Adherent cells Wash twice with PBS, add trypsin to digest the cells, and centrifuge at 1000g for 3-5 minutes after digestion is completed.
(3) Add 1mL of DAPI working solution to resuspend the cells and incubate at room temperature in the dark for 5-30 minutes. The optimal incubation time for different cells is different, please explore by yourself according to the specific experimental needs.
(4) After the incubation, centrifuge at 1000g for 5 minutes, remove the supernatant, add PBS and wash 2-3 times, 5 minutes each time.
(5) Resuspend the cells in pre-warmed serum-free cell culture medium or PBS. Observe by fluorescence microscopy or flow cytometry.
3. Cell adhesion staining
(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 dye working solution from one corner of the coverslip and shake gently to evenly cover all cells with the dye.
(4) Incubate at room temperature in the dark for 3-5 minutes. The optimal incubation time for different cells is different, please explore by yourself according to the specific experimental needs.
(5) After the incubation, discard the dye working solution and use pre-warmed culture solution to wash the coverslip 2 to 3 times.
4. Microscope detection The maximum excitationemission light of DAPI is 364454nm respectively.
Precautions
① DAPI is suitable for staining fixed cells or tissues, usually after other staining;
② To slow down fluorescence quenching, anti-fluorescence quenching mounting agent can be used;
③ Fluorescent dyes all have quenching problems. Please try to avoid light to slow down fluorescence quenching;
④ DAPI is irritating to the human body. For your safety and health, please wear a lab coat and disposable gloves when operating.
| Cas No. | 28718-90-3 | SDF | |
| Synonyms | 4',6Diamidino2phenylindole, FxCycle Violet | ||
| Canonical SMILES | NC(C(C=C1)=CC=C1C2=CC3=CC=C(C(N)=N)C=C3N2)=N.Cl.Cl | ||
| Formula | C16H15N5•2HCl | M.Wt | 350.3 |
| Solubility | Insoluble in EtOH; ≥10 mg/mL in Water; ≥53.3 mg/mL in DMSO | Storage | Store at -20°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 | 2.8547 mL | 14.2735 mL | 28.547 mL |
| 5 mM | 570.9 μL | 2.8547 mL | 5.7094 mL |
| 10 mM | 285.5 μL | 1.4273 mL | 2.8547 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
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Average Rating: 5 (Based on Reviews and 32 reference(s) in Google Scholar.)