DAPI [4,6-Diamidino-2-phenylindole, dihydrochloride] 10 mM solution in water

$L. pneumophila infection increases the secretion of EVs in THP-1 cells. (a) Amount of EVs in response to L. pneumophila infection. THP-1 cells were treated with IL-1β (1 ng/mL) or infected with L. pneumophila (MOI 0.25 or 0.5, respectively) for 24 h. NTA was performed with the distinct particle fractions separated by differential centrifugation. (b) Western blot for exosomal marker proteins. Whole cell lysate or 100 k pellet derived from uninfected THP-1 cells were used. Equal protein amounts were loaded. (c) Transmission electron microscopy with 100 k pellet. Purified EVs from THP-1 cells were fixed and visualized after negative staining with uranyl acetate. Scale bar represents 100 nm. (d) Uptake of A549-derived EVs by THP-1 cells. A549 cells were stained with the membrane dye PKH67. EVs were collected (100 k pellet) and incubated with THP-1 cells for 1 or 3 h, respectively. Nuclei were stained by DAPI and pictures were taken with an original magnification of 630x. A: Data are shown as mean + SEM of three independent experiments. **p < 0.01, ***p < 0.001, ****p < 0.0001. B–D: representative results. Source: Legionella pneumophila infection activates bystander cells differentially by bacterial and host cell vesicles by Jung et al., Scientific Reports, July 2017.$

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Physical properties

Molecular weight	350.25
Solvent	Water

Spectral properties

Extinction coefficient (cm ^-1 M ^-1)	27000
Excitation (nm)	359
Emission (nm)	457

Storage, safety and handling

Certificate of Origin	Download PDF
H-phrase	H303, H313, H340
Hazard symbol	T
Intended use	Research Use Only (RUO)
R-phrase	R20, R21, R68
Storage	Freeze (< -15 °C); Minimize light exposure
UNSPSC	41116134

Alternative formats

DAPI [4,6-Diamidino-2-phenylindole, dihydrochloride] *CAS 28718-90-3*

Overview SDS Protocol

CAS

28718-90-3

Molecular weight

350.25

Extinction coefficient (cm ^-1 M ^-1)

27000

Excitation (nm)

359

Emission (nm)

457

DAPI is a fluorescent stain that binds strongly to DNA. It is used extensively in fluorescence microscopy. Since DAPI passes through an intact cell membrane, it can be used to stain live cells besides fixed cells. For fluorescence microscopy, DAPI is excited with ultraviolet light. When bound to double-stranded DNA its absorption maximum is at 358 nm and its emission maximum is at 461 nm. One drawback of DAPI is that its emission is fairly broad. DAPI also binds to RNA although it is not as strongly fluorescent as it binds to DNA. Its emission shifts to around 500 nm when bound to RNA. DAPI's blue emission is convenient for multiplexing assays since there is very little fluorescence overlap between DAPI and green-fluorescent molecules like fluorescein and green fluorescent protein (GFP), or red-fluorescent stains like Texas Red. Besides labeling cell nuclei, DAPI is also used for the detection of mycoplasma or virus DNA in cell cultures.

Calculators

Common stock solution preparation

Table 1. Volume of Water needed to reconstitute specific mass of DAPI [4,6-Diamidino-2-phenylindole, dihydrochloride] *10 mM solution in water* to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

	0.1 mg	0.5 mg	1 mg	5 mg	10 mg
1 mM	285.51 µL	1.428 mL	2.855 mL	14.276 mL	28.551 mL
5 mM	57.102 µL	285.51 µL	571.021 µL	2.855 mL	5.71 mL
10 mM	28.551 µL	142.755 µL	285.51 µL	1.428 mL	2.855 mL

Molarity calculator

Enter any two values (mass, volume, concentration) to calculate the third.

Mass (Calculate)		Molecular weight		Volume (Calculate)		Concentration (Calculate)		Moles
	/		=		x		=

Spectrum

Open in Advanced Spectrum Viewer

Spectral properties

Extinction coefficient (cm ^-1 M ^-1)	27000
Excitation (nm)	359
Emission (nm)	457

Images

Figure 1. L. pneumophila infection increases the secretion of EVs in THP-1 cells. (a) Amount of EVs in response to L. pneumophila infection. THP-1 cells were treated with IL-1β (1 ng/mL) or infected with L. pneumophila (MOI 0.25 or 0.5, respectively) for 24 h. NTA was performed with the distinct particle fractions separated by differential centrifugation. (b) Western blot for exosomal marker proteins. Whole cell lysate or 100 k pellet derived from uninfected THP-1 cells were used. Equal protein amounts were loaded. (c) Transmission electron microscopy with 100 k pellet. Purified EVs from THP-1 cells were fixed and visualized after negative staining with uranyl acetate. Scale bar represents 100 nm. (d) Uptake of A549-derived EVs by THP-1 cells. A549 cells were stained with the membrane dye PKH67. EVs were collected (100 k pellet) and incubated with THP-1 cells for 1 or 3 h, respectively. Nuclei were stained by DAPI and pictures were taken with an original magnification of 630x. A: Data are shown as mean + SEM of three independent experiments. **p < 0.01, ***p < 0.001, ****p < 0.0001. B–D: representative results. Source: Legionella pneumophila infection activates bystander cells differentially by bacterial and host cell vesicles by Jung et al., Scientific Reports, July 2017.

Figure 2. Chemical structure for DAPI [4,6-Diamidino-2-phenylindole, dihydrochloride] *10 mM solution in water*

Citations

View all 40 citations: Citation Explorer

Metabolic programming determines the pathogenicity of Campylobacter jejuni
Authors: Hosomi, Koji and Hatanaka, Noritoshi and Hinenoya, Atsushi and Tojima, Yoko and Furuta, Mari and Nagatake, Takahiro and Saika, Azusa and Kawai, Soichiro and Yoshii, Ken and Kondo, Saki and others,
Journal: (2023)

The sage genome provides insight into the evolutionary dynamics of diterpene biosynthesis gene cluster in plants
Authors: Li, Chen-Yi and Yang, Lei and Liu, Yan and Xu, Zhou-Geng and Gao, Jian and Huang, Yan-Bo and Xu, Jing-Jing and Fan, Hang and Kong, Yu and Wei, Yu-Kun and others,
Journal: Cell Reports (2022): 111236

Adipose-Derived Stem Cell-Incubated HA-Rich Sponge Matrix Implant Modulates Oxidative Stress to Enhance VEGF and TGF-$\beta$ Secretions for Extracellular Matrix Reconstruction In Vivo
Authors: Cheng, Yu-Shen and Yen, Hung-Hsun and Chang, Chung-Yen and Lien, Wei-Chih and Huang, Shu-Hung and Lee, Su-Shin and Wang, Lin and Wang, Hui-Min David
Journal: Oxidative Medicine and Cellular Longevity (2022)

Carboxyl-Terminal Decoy Epitopes in the Capsid Protein of Porcine Circovirus Type 2 Are Immunogenicity-Enhancers That Elicit Predominantly Specific Antibodies in Non-Vaccinated Pigs
Authors: Hung, Ling-Chu
Journal: Viruses (2022): 2373

Modulating Microglia/Macrophage Activation by CDNF Promotes Transplantation of Fetal Ventral Mesencephalic Graft Survival and Function in a Hemiparkinsonian Rat Model
Authors: Tseng, Kuan-Yin and Wu, Jui-Sheng and Chen, Yuan-Hao and Airavaara, Mikko and Cheng, Cheng-Yi and Ma, Kuo-Hsing
Journal: Biomedicines (2022): 1446

DLP 3D printed silica-doped HAp ceramic scaffolds inspired by the trabecular bone structure
Authors: Zhang, Chengyu and Yuan, Yanping and Zeng, Yong and Chen, Jimin
Journal: Ceramics International (2022): 27765--27773

Assessment of Cellular Alignment and Proliferation on Cellulose Nanofibril (CNF) Films
Authors: Roozbahani, Sahar
Journal: (2022)

The Autophagy Receptor TAX1BP1 (T6BP) improves antigen presentation by MHC-II molecules
Authors: Sarango, Gabriela and Richetta, Cl{\'e}mence and Pereira, Mathias and Kumari, Anita and Ghosh, Michael and Bertrand, Lisa and Pionneau, C{\'e}dric and Le Gall, Morgane and Gr{\'e}goire, Sylvie and Jeger-Madiot, Rapha{\"e}l and others,
Journal: EMBO reports (2022): e55470

Novel SPEF2 Variant in a Japanese Patient with Primary Ciliary Dyskinesia: A Case Report and Literature Review
Authors: Mori, Mayako and Kido, Takashi and Sakamoto, Noriho and Ozasa, Mutsumi and Kido, Kumiko and Noguchi, Yasuko and Tokito, Takatomo and Okuno, Daisuke and Yura, Hirokazu and Hara, Atsuko and others,
Journal: Journal of Clinical Medicine (2022): 317

A robust mechanism for resetting juvenility during each generation in Arabidopsis
Authors: Gao, Jian and Zhang, Ke and Cheng, Ying-Juan and Yu, Sha and Shang, Guan-Dong and Wang, Fu-Xiang and Wu, Lian-Yu and Xu, Zhou-Geng and Mai, Yan-Xia and Zhao, Xin-Yan and others,
Journal: Nature Plants (2022): 257--268

References

View all 134 references: Citation Explorer

Reliable DNA ploidy determination in dehydrated tissues of vascular plants by DAPI flow cytometry--new prospects for plant research
Authors: Suda J, Travnicek P.
Journal: Cytometry A (2006): 273

Identification of all pachytene bivalents in the common shrew using DAPI-staining of synaptonemal complex spreads
Authors: Belonogova NM, Karamysheva TV, Biltueva LS, Perepelov EA, Minina JM, Polyakov AV, Zhdanova NS, Rubtsov NB, Searle JB, Borodin PM.
Journal: Chromosome Res (2006): 673

Physical localization of ribosomal genes and chromosome DAPI banding by in situ hybridization in Medicago sativa L
Authors: Chen JM, Hong YH, Wang YP, Bowley S, Wan JM.
Journal: Yi Chuan (2006): 184

DNA staining with the fluorochromes EtBr, DAPI and YOYO-1 in the comet assay with tobacco plants after treatment with ethyl methanesulphonate, hyperthermia and DNase-I
Authors: Gichner T, Mukherjee A, Veleminsky J.
Journal: Mutat Res (2006): 17

Investigation of chromosomes in varieties and translocation lines of pea Pisum sativum L. by FISH, Ag-NOR, and differential DAPI staining
Authors: Samatadze TE, Muravenko OM, Bol'sheva NL, Amosova AB, Gostimsckii SA, Zelenin AV.
Journal: Genetika (2005): 1665

DAPI fluorescence in nuclei isolated from tumors
Authors: Krishan A, D and ekar PD., undefined
Journal: J Histochem Cytochem (2005): 1033

Polarized fluorescence correlation spectroscopy of DNA-DAPI complexes
Authors: Barcellona ML, Gammon S, Hazlett T, Digman MA, Gratton E.
Journal: Microsc Res Tech (2004): 205

DNA quantification in nuclei of cultivated mushroom with DAPI staining
Authors: Pancheva EV, Volkova VN, Kamzolkina OV.
Journal: Tsitologiia (2004): 381

Quantification of sPLA2-induced early and late apoptosis changes in neuronal cell cultures using combined TUNEL and DAPI staining
Authors: Daniel B, DeCoster MA.
Journal: Brain Res Brain Res Protoc (2004): 144

DAPI derivative: a fluorescent DNA dye that can be covalently attached to biomolecules
Authors: Li M, Wu RS, Tsai JS.
Journal: Bioorg Med Chem Lett (2003): 4351