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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-1β (1 ng/mL) or infected with <em>L. pneumophila</em> (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: <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-1β (1 ng/mL) or infected with <em>L. pneumophila</em> (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: <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.
Chemical structure for DAPI [4,6-Diamidino-2-phenylindole, dihydrochloride] *10 mM solution in water*
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Catalog Number17507
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Telephone1-408-733-1055
Fax1-408-733-1304
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ShippingStandard overnight for United States, inquire for international
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
Alternative formats
DAPI [4,6-Diamidino-2-phenylindole, dihydrochloride] *CAS 28718-90-3*

OverviewpdfSDSpdfProtocol


See also: Nucleus, Bioconjugation, Cell Cycle Assays, DNA and RNA Quantitation
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

Molarity calculator

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

Mass (Calculate)Molecular weightVolume (Calculate)Concentration (Calculate)Moles
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Spectrum


Open in Advanced Spectrum Viewer
spectrum

Spectral properties

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

Images


<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-1β (1 ng/mL) or infected with <em>L. pneumophila</em> (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: <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.
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.
Chemical structure for DAPI [4,6-Diamidino-2-phenylindole, dihydrochloride] *10 mM solution in water*
Figure 2. Chemical structure for DAPI [4,6-Diamidino-2-phenylindole, dihydrochloride] *10 mM solution in water*

Citations


View all 26 citations: Citation Explorer
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)
Dihydromyricetin suppresses cell metastasis in human osteosarcoma through SP-1-and NF-$\kappa$B-modulated urokinase plasminogen activator inhibition
Authors: Chou, Chia-Hsuan and Lu, Ko-Hsiu and Yang, Jia-Sin and Hsieh, Yi-Hsien and Lin, Chiao-Wen and Yang, Shun-Fa
Journal: Phytomedicine (2021): 153642
A Novel Dibenzoxazepine Attenuates Intracellular Salmonella Typhimurium Oxidative Stress Resistance
Authors: Hsu, Cheng-Yun and Wu, Yi-Lun and Lin, Hsueh-Chun and Lin, Man-Yi and Chou, Shih-Hsiu and Shiau, Chung-Wai and Chiu, Hao-Chieh
Journal: Microbiology spectrum (2021): e01519--21
Thermo/photo dual-crosslinking chitosan-gelatin methacrylate hydrogel with controlled shrinking property for contraction fabrication
Authors: Luo, Xin and Liu, Yidan and Pang, Jianhui and Bi, Shichao and Zhou, Zhongzheng and Lu, Zhe and Feng, Chao and Chen, Xiguang and Kong, Ming
Journal: Carbohydrate Polymers (2020): 116067
Noncoding RNA MaIL1 is an integral component of the TLR4--TRIF pathway
Authors: Aznaourova, Marina and Janga, Harshavardhan and Sefried, Stephanie and Kaufmann, Andreas and Dorna, Jens and Volkers, Sarah M and Georg, Philipp and Lechner, Marcus and Hoppe, Judith and D{\"o}kel, Simon and others,
Journal: Proceedings of the National Academy of Sciences (2020): 9042--9053
Akt regulates a Rab11-effector switch required for ciliogenesis
Authors: Walia, Vijay and Cuenca, Adrian and Vetter, Melanie and Insinna, Christine and Perera, Sumeth and Lu, Quanlong and Ritt, Daniel A and Semler, Elizabeth and Specht, Suzanne and Stauffer, Jimmy and others,
Journal: Developmental cell (2019): 229--246
Immunological association of inducible bronchus-associated lymphoid tissue organogenesis in Ag85B-rHPIV2 vaccine-induced anti-tuberculosis mucosal immune responses in mice
Authors: Nagatake, Takahiro and Suzuki, Hidehiko and Hirata, So-ichiro and Matsumoto, Naomi and Wada, Yasuko and Morimoto, Sakiko and Nasu, Ayaka and Shimojou, Michiko and Kawano, Mitsuo and Ogami, Kentaro and others,
Journal: International immunology (2018): 471--481
Astrocytic expression of the CXCL12 receptor, CXCR7/ACKR3 is a hallmark of the diseased, but not developing CNS
Authors: Puchert, Malte and Pelkner, Fabian and Stein, Gregor and Angelov, Doychin N and Boltze, Johannes and Wagner, Daniel-Christoph and Odoardi, Francesca and Flügel, Alex and er , undefined and Streit, Wolfgang J and Engele, Jürgen
Journal: Molecular and Cellular Neuroscience (2017)
Legionella pneumophila infection activates bystander cells differentially by bacterial and host cell vesicles
Authors: Jung, Anna Lena and Herkt, Christina Elena and Schulz, Christine and Bolte, Kathrin and Seidel, Kerstin and Scheller, Nicoletta and Sittka-Stark, Alexandra and Bertrams, Wilhelm and Schmeck, Bernd
Journal: Scientific reports (2017): 1--11
Peptides mimicking viral proteins of porcine circovirus type 2 were profiled by the spectrum of mouse anti-PCV2 antibodies
Authors: Hung, Ling-Chu and Yang, Cheng-Yao and Cheng, Ivan-Chen
Journal: BMC immunology (2017): 25

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

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Certificate of origin
Safety data sheet
Product protocol
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Application notes


10 Best DNA Probes
Selective Analysis of RNA in Live and Fixed Cells with StrandBrite RNA Green

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