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

<em>L. pneumophila</em> infection increases the secretion of EVs in THP-1 cells. (a) Amount of EVs in response to <em>L. pneumophila</em> infection. THP-1 cells were treated with IL-1&beta; (1&thinsp;ng/mL) or infected with <em>L. pneumophila</em> (MOI 0.25 or 0.5, respectively) for 24&thinsp;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&thinsp;k pellet derived from uninfected THP-1 cells were used. Equal protein amounts were loaded. (c) Transmission electron microscopy with 100&thinsp;k pellet. Purified EVs from THP-1 cells were fixed and visualized after negative staining with uranyl acetate. Scale bar represents 100&thinsp;nm. (d) Uptake of A549-derived EVs by THP-1 cells. A549 cells were stained with the membrane dye PKH67. EVs were collected (100&thinsp;k pellet) and incubated with THP-1 cells for 1 or 3&thinsp;h, respectively. Nuclei were stained by DAPI and pictures were taken with an original magnification of 630x. A: Data are shown as mean&thinsp;+&thinsp;SEM of three independent experiments. **p&thinsp;&lt;&thinsp;0.01, ***p&thinsp;&lt;&thinsp;0.001, ****p&thinsp;&lt;&thinsp;0.0001. B&ndash;D: representative results. Source: <strong><em>Legionella pneumophila</em> infection activates bystander cells differentially by bacterial and host cell vesicles </strong>by Jung et al., <em>Scientific Reports,</em> July 2017.
<em>L. pneumophila</em> infection increases the secretion of EVs in THP-1 cells. (a) Amount of EVs in response to <em>L. pneumophila</em> infection. THP-1 cells were treated with IL-1&beta; (1&thinsp;ng/mL) or infected with <em>L. pneumophila</em> (MOI 0.25 or 0.5, respectively) for 24&thinsp;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&thinsp;k pellet derived from uninfected THP-1 cells were used. Equal protein amounts were loaded. (c) Transmission electron microscopy with 100&thinsp;k pellet. Purified EVs from THP-1 cells were fixed and visualized after negative staining with uranyl acetate. Scale bar represents 100&thinsp;nm. (d) Uptake of A549-derived EVs by THP-1 cells. A549 cells were stained with the membrane dye PKH67. EVs were collected (100&thinsp;k pellet) and incubated with THP-1 cells for 1 or 3&thinsp;h, respectively. Nuclei were stained by DAPI and pictures were taken with an original magnification of 630x. A: Data are shown as mean&thinsp;+&thinsp;SEM of three independent experiments. **p&thinsp;&lt;&thinsp;0.01, ***p&thinsp;&lt;&thinsp;0.001, ****p&thinsp;&lt;&thinsp;0.0001. B&ndash;D: representative results. Source: <strong><em>Legionella pneumophila</em> infection activates bystander cells differentially by bacterial and host cell vesicles </strong>by Jung et al., <em>Scientific Reports,</em> July 2017.
<em>L. pneumophila</em> infection increases the secretion of EVs in THP-1 cells. (a) Amount of EVs in response to <em>L. pneumophila</em> infection. THP-1 cells were treated with IL-1&beta; (1&thinsp;ng/mL) or infected with <em>L. pneumophila</em> (MOI 0.25 or 0.5, respectively) for 24&thinsp;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&thinsp;k pellet derived from uninfected THP-1 cells were used. Equal protein amounts were loaded. (c) Transmission electron microscopy with 100&thinsp;k pellet. Purified EVs from THP-1 cells were fixed and visualized after negative staining with uranyl acetate. Scale bar represents 100&thinsp;nm. (d) Uptake of A549-derived EVs by THP-1 cells. A549 cells were stained with the membrane dye PKH67. EVs were collected (100&thinsp;k pellet) and incubated with THP-1 cells for 1 or 3&thinsp;h, respectively. Nuclei were stained by DAPI and pictures were taken with an original magnification of 630x. A: Data are shown as mean&thinsp;+&thinsp;SEM of three independent experiments. **p&thinsp;&lt;&thinsp;0.01, ***p&thinsp;&lt;&thinsp;0.001, ****p&thinsp;&lt;&thinsp;0.0001. B&ndash;D: representative results. Source: <strong><em>Legionella pneumophila</em> infection activates bystander cells differentially by bacterial and host cell vesicles </strong>by Jung et al., <em>Scientific Reports,</em> July 2017.
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Physical properties
Molecular weight350.25
SolventWater
Spectral properties
Extinction coefficient (cm -1 M -1)27000
Excitation (nm)359
Emission (nm)457
Storage, safety and handling
Certificate of OriginDownload PDF
H-phraseH303, H313, H340
Hazard symbolT
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R68
StorageFreeze (< -15 °C); Minimize light exposure
UNSPSC41116134

OverviewpdfSDSpdfProtocol


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 mg0.5 mg1 mg5 mg10 mg
1 mM285.51 µL1.428 mL2.855 mL14.276 mL28.551 mL
5 mM57.102 µL285.51 µL571.021 µL2.855 mL5.71 mL
10 mM28.551 µL142.755 µL285.51 µL1.428 mL2.855 mL

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Spectrum


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Spectral properties

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

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Citations


View all 47 citations: Citation Explorer
Mechanosensitive membrane domains regulate calcium entry in arterial endothelial cells to protect against inflammation
Authors: Hong, Soon-Gook and Ashby, Julianne W and Kennelly, John P and Wu, Meigan and Steel, Michelle and Chattopadhyay, Eesha and Foreman, Rob and Tontonoz, Peter and Tarling, Elizabeth J and Turowski, Patric and others,
Journal: The Journal of Clinical Investigation (2024)
A novel HDAC8 inhibitor H7E exerts retinoprotective effects against glaucomatous injury via ameliorating aberrant M{\"u}ller glia activation and oxidative stress
Authors: Wu, Liang-Huan and Cheng, Yu-Wen and Lin, Fan-Li and Hsu, Kai-Cheng and Wang, Mong-Heng and Yen, Jing-Lun and Wang, Tsung-Jen and Lin, Tony Eight and Liu, Yi-Chien and Huang, Wei-Jan and others,
Journal: Biomedicine \& Pharmacotherapy (2024): 116538
A xenotransplantation mouse model to study physiology of the mammary gland from large mammals
Authors: Miller, James L and Reddy, Alexandra and Harman, Rebecca M and Van de Walle, Gerlinde R
Journal: Plos one (2024): e0298390
Co-Delivery of Bioengineered Exosomes and Oxygen for Treating Critical Limb Ischemia in Diabetic Mice
Authors: Zhong, Ting and Gao, Ning and Guan, Ya and Liu, Zhongting and Guan, Jianjun
Journal: ACS nano (2023)
Alcaligene s Lipid A Functions as a Superior Mucosal Adjuvant to Monophosphoryl Lipid A via the Recruitment and Activation of CD11b+ Dendritic Cells in Nasal Tissue
Authors: Sun, Xiao and Hosomi, Koji and Shimoyama, Atsushi and Yoshii, Ken and Saika, Azusa and Yamaura, Haruki and Nagatake, Takahiro and Kiyono, Hiroshi and Fukase, Koichi and Kunisawa, Jun
Journal: International Immunology (2023): dxad045
Bio-AFM exploits enhanced response of human gingival fibroblasts on TiO2 nanotubular substrates with thin TiO2 coatings
Authors: Baishya, Kaushik and Vrchoveck{\'a}, Kate{\v{r}}ina and Alijani, Mahnaz and Rodriguez-Pereira, Jhonatan and Thalluri, Sitaramanjaneya Mouli and Goldbergov{\'a}, Monika P{\'a}vkov{\'a} and P{\v{r}}ibyl, Jan and Macak, Jan M
Journal: Applied Surface Science Advances (2023): 100459
Cerebral Myelination in a Bronchopulmonary Dysplasia Murine Model
Authors: Chen, Wenwen and Wang, Ran and Chen, Chao
Journal: Children (2023): 1321
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)
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

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