DAPI
4,6-Diamidino-2-phenylindole, dihydrochloride; CAS 28718-90-3
DAPI (4'-6-diamidino-2-phenylindole) is a DNA-specific blue fluorescent dye used in many biological disciplines, including biochemistry, molecular biology, immunology, and microbiology. Because of its 20-fold fluorescence increase and high binding affinity for adenine-thymine (AT) rich regions of double-stranded DNA (dsDNA), DAPI is an excellent dye for cell counting and sorting, measuring apoptosis, and as a nuclear counterstain in fluorescence microscopy and flow cytometry. While DAPI is more sensitive for use with dsDNA, it also binds to adenine-uracil (AU) regions of RNA but with less affinity and fluorescence enhancement.
When bound to dsDNA, DAPI has an excitation/emission maxima of 359/457 nm. It is well-excited by the violet (405 nm) laser and detected through the DAPI filter. The high quantum yield (Φ = 0.92) of the DAPI/dsDNA complex enables it to emit bright blue fluorescence making DAPI an excellent counterstain in multicolor fluorescent techniques with other green (e.g., iFluor® 488, Alexa Fluor® 488, FITC, GFP) and red (e.g., iFluor® 647, Alexa Fluor® 647, Cy5, RFP) fluorophores. DAPI will also bind to RNA but at a lower affinity and emit a significantly weaker fluorescence signal, ∼20% as high as the yield of the DAPI/dsDNA complex. The DAPI/RNA complex also exhibits a longer-wavelength emission maximum at ∼500 nm.
In most applications, cell/tissue fixation and permeabilization are done before DAPI staining since the dye is moderately cell impermeant. Increasing the DAPI concentration or incubation time may be necessary. However, this could lead to decreased cell viability. For live-cell DNA staining, Nuclear Violet™ LCS1 and Hoeschts dyes are popular alternatives. They have absorption and fluorescence emission maxima similar to DAPI and are cell-permeant. Besides labeling cell nuclei, DAPI is also used to detect mycoplasma or virus DNA in cell cultures.
In most applications, cell/tissue fixation and permeabilization are done before DAPI staining since the dye is moderately cell impermeant. Increasing the DAPI concentration or incubation time may be necessary. However, this could lead to decreased cell viability. For live-cell DNA staining, Nuclear Violet™ LCS1 and Hoeschts dyes are popular alternatives. They have absorption and fluorescence emission maxima similar to DAPI and are cell-permeant. Besides labeling cell nuclei, DAPI is also used to detect 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] *CAS 28718-90-3* 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
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Spectrum
Citations
View all 87 citations: Citation Explorer
An NIR-responsive hydrogel loaded with polydeoxyribonucleotide nano-vectors for enhanced chronic wound healing
Authors: Sun, Yanzhen and Li, Yao and Ding, Xiaokang and Xu, Pan and Jing, Xiaodong and Cong, Hailin and Hu, Hao and Yu, Bing and Xu, Fu-Jian
Journal: Biomaterials (2024): 122789
Authors: Sun, Yanzhen and Li, Yao and Ding, Xiaokang and Xu, Pan and Jing, Xiaodong and Cong, Hailin and Hu, Hao and Yu, Bing and Xu, Fu-Jian
Journal: Biomaterials (2024): 122789
Emergence of large-scale cell death through ferroptotic trigger waves
Authors: Co, Hannah KC and Wu, Chia-Chou and Lee, Yi-Chen and Chen, Sheng-hong
Journal: Nature (2024): 1--9
Authors: Co, Hannah KC and Wu, Chia-Chou and Lee, Yi-Chen and Chen, Sheng-hong
Journal: Nature (2024): 1--9
Deciphering the Role of Ferroptosis in Rheumatoid Arthritis: Synovial Transcriptome Analysis and Immune Infiltration Correlation
Authors: Wang, Hongli and Zhang, Miaomiao and Hu, Yiping and He, Juan and Zhong, Yuchao and Dai, Yong and Wang, Qingwen
Journal: Heliyon (2024)
Authors: Wang, Hongli and Zhang, Miaomiao and Hu, Yiping and He, Juan and Zhong, Yuchao and Dai, Yong and Wang, Qingwen
Journal: Heliyon (2024)
The mesenchymal stromal cell (MSC) secretome promotes tissue regeneration and increases macrophage infiltration in acute and MRSA-infected skin wounds in vivo
Authors: Rajesh, Aarthi and Ju, Esther Da Eun and Oxford, Kelly A and Harman, Rebecca M and Van de Walle, Gerlinde R
Journal: Cytotherapy (2024)
Authors: Rajesh, Aarthi and Ju, Esther Da Eun and Oxford, Kelly A and Harman, Rebecca M and Van de Walle, Gerlinde R
Journal: Cytotherapy (2024)
Adipose Tissue and Translation Machinery in Metabolic Regulation
Authors: De Siqueira, Mirian Krystel
Journal: (2024)
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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
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
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
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
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
Authors: Samatadze TE, Muravenko OM, Bol'sheva NL, Amosova AB, Gostimsckii SA, Zelenin AV.
Journal: Genetika (2005): 1665
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