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DAPI (4',6-Diamidino-2-Phenylindole, Dilactate) DAPI (4',6-Diamidino-2-Phenylindole, Dilactate)

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Regulatory Status
RUO
Other Names
4',6-Diamidino-2-Phenylindole, Dilactate
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Product Citations
publications
1_Ankita_Patil.jpg
Methanol fixed superior cervical ganglion neurons were stained with anti-tubulin β3 (pink) and anti-TRIM46 (aqua), then stained with secondary antibodies, phalloidin (orange) and DAPI (blue). Image generously submitted to the 2017 Cell Life Imaging Competition by Ankita Patil from Drexel University.
  • 1_Ankita_Patil.jpg
    Methanol fixed superior cervical ganglion neurons were stained with anti-tubulin β3 (pink) and anti-TRIM46 (aqua), then stained with secondary antibodies, phalloidin (orange) and DAPI (blue). Image generously submitted to the 2017 Cell Life Imaging Competition by Ankita Patil from Drexel University.
  • 2_Kathleen-Ignatoski
    Frozen mouse intestine stained with purified anti-Ankyrin-B (green), E-Cadherin (red) and DAPI (blue) followed by anti-goat Alexa Fluor® 488 and anti-rabbit Alexa Fluor® 568. Image generously submitted to the 2017 Cell Life Imaging Competition by Kathleen Ignatoski from University of Michigan.
  • 3_KifLiakath.jpg
    Whole mount mouse tail epidermis was stained with purified anti-Keratin 14 (green, clone Poly19053), purified anti-Keratin 15 (orange, clone Poly18339) and DAPI (blue). Image generously submitted to the 2017 Cell Life Imaging Competition by Kif Liakath.
  • 4_DAPI_IF_080712
    Human cervical cancer cell line, HeLa, was fixed with 4% paraformaldehyde and stained with 300 nM DAPI.
  • 5_DAPI_080712
    Human cervical cancer cell line, HeLa, was fixed with 4% paraformaldehyde and stained with 300 nM DAPI.
See DAPI (4',6-Diamidino-2-Phenylindole, Dilactate) spectral data
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422801 10 mg DKK600
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Description

DAPI (4',6-Diamidino-2-Phenylindole, Dilactate), a blue fluorescent nucleic acid stain, preferentially binds dsDNA and associates with the minor groove AT clusters. The molecular weight for DAPI is 457.5.  Association of DAPI with dsDNA increases fluorescence approximately 20-fold, with an emission maximum of 460 nm. The increase in fluorescence is believed to be due to displacement of water molecules from the minor groove and DAPI. DAPI can also bind RNA and emits at a longer wavelength of 500 nm, with only a 20% increase in fluorescence.

DAPI is a popular nuclear stain for identification of cell cycle and specifically stains nuclei but not cytoplasm. The dilactate form is more water soluble compared to the dihydrochloride form.

DAPI can be excited by a UV laser, xenon, or a mercury-arc lamp. 

Product Details
Technical Data Sheet (pdf)

Product Details

Formulation
Lyophilized
Preparation
To make 5 mg/ml (10.9 mM) solution, dissolve entire contents of vial in 2 ml of deionized water.
Storage & Handling
Store between 2°C and 8°C, protected from light. For long term storage, aliquot and store at -20°C.
Application

ICFC, ICC
Recommended Usage

This product can be used at 30 µM for flow cytometry and 300 nM for immunofluorescence.  It is recommended to titrate the dye for optimal performance. 

Excitation Laser
Ultraviolet Laser (355 nm)
Application Notes

Protocols for Counterstaining with DAPI:

Immunofluorescence Staining
Note: Cells may be fixed by method of choice, such as with 4% paraformaldehyde. DAPI staining is done after staining for other markers. Fixation and permeabilization of cells is not necessary to counterstain with DAPI.

1. Fix cells using method of choice.
2. Incubate the cells with phosphate buffered saline (PBS) for 15 minutes.
3. Dilute the DAPI solution (we recommend to 300 nM) with PBS. Remove excess PBS from the slide and cover with DAPI solution, making sure the cells are completely covered.
4. Incubate for up to 5 minutes. Rinse the slide several times to remove all free DAPI. It is recommended to use a mounting medium with antifade reagent to reduce fluorescence quenching. The slide is ready to view under the microscope with appropriate filters.


Nuclear Staining for Flow Cytometry
Note: Cells may be fixed with 4% paraformadehyde or absolute ethanol.

1. Fix cells with absolute ethanol or 4% paraformaldehyde.
2. Centrifuge, discard the supernatant, and add 5 ml of PBS, allowing cells to rehydrate for 15 minutes.
3. Dilute the DAPI to 30 µM (recommended) in staining buffer (such as BioLegend's FOXP3 Perm Buffer, Cat. No. 421402).
4. Centrifuge cell suspension from step 3 above, discard supernatant, and add 1 ml of DAPI solution; then incubate at room temperature for 15 minutes.
5. Analyze by flow cytometry using an instrument with appropriate lasers.

Additional Product Notes

View more applications data for this product in our Scientific Poster Library.

Application References
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  208. Zhang M, et al. 2017. Oncol Lett. 3.069444444. PubMed
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  216. DeDreu J, et al. 2020. FASEB J. 34:9316. PubMed
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  218. Deng J, et al. 2021. Breast Cancer Res Treat. 185:85. PubMed
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  220. Kanda K, et al. 2018. JCI Insight. 3:e91316. PubMed
  221. Fujiwara H, et al. 2021. Nat Immunol. 22:1440. PubMed
  222. Ravi A, et al. 2021. Dev Cell. 56:1661. PubMed
  223. Wu CL, et al. 2021. Nat Commun. 12:362. PubMed
  224. Koh P, et al. 2016. Sci Data. 3:160109. PubMed
  225. Wang ECE, et al. 2019. Cell Stem Cell. 24:654. PubMed
  226. Poulos M, et al. 2016. Nat Commun. 7:13829. PubMed
  227. Noah AC, et al. 2020. J Appl Physiol (1985). 473:128. PubMed
  228. So EY, et al. 2021. Am J Physiol Cell Physiol. 321:C569. PubMed
  229. Teo YV, et al. 2019. Cell Rep. 27:997. PubMed
  230. Hayama Y, et al. 2018. J Immunol. 200:3790. PubMed
  231. Fredriksson-Lidman K, et al. 2017. PLoS One. 10.1371/journal.pone.0185509. PubMed
  232. Wennerberg E, et al. 2022. Sci Transl Med. 14:eabe8195. PubMed
  233. Kumar A, et al. 2021. STAR Protoc. 2:100874. PubMed
  234. Hendricks A, et al. 2020. Cancers (Basel). 12:00. PubMed
  235. Aronovich A, et al. 2021. J Invest Dermatol. 141:619. PubMed
  236. Faisal S, et al. 2016. Sci Rep. 6:39530. PubMed
  237. Caeser R, et al. 2021. Nat Protoc. 16:2499. PubMed
  238. Thompson M, et al. 2015. J Immunol. 195: 5637 - 5647. PubMed
  239. Kundu K, et al. 2019. Cancer Immunol Res. 1.069444444. PubMed
  240. Pagan JD, et al. 2018. Cell. 172:564. PubMed
  241. Zhang X, et al. 2022. JCI Insight. 7:. PubMed
  242. Zhu M, et al. 2021. Eneuro. 8:. PubMed
  243. Flores-Pliego A, et al. 2021. Cells. 10:. PubMed
  244. Shani O, et al. 2021. Elife. 10: . PubMed
  245. Mould KJ, et al. 2019. JCI Insight. 4:00. PubMed
  246. Moser J, et al. 2016. Sci Rep. 6:35387. PubMed
  247. Heath O, et al. 2021. Cancer Immunol Res. 9:665. PubMed
  248. Dasinger JH, et al. 2021. Pregnancy Hypertens. 24:126. PubMed
  249. Ren X, et al. 2022. STAR Protoc. 3:101818. PubMed
  250. Correa A, et al. 2015. PLoS One. 10: 0143391. PubMed
  251. Yanting C, et al. 2018. J Dairy Sci. 101:1601. PubMed
  252. Kurena B, et al. 2017. Front Immunol. 1.490972222. PubMed
  253. Pinho–Ribeiro FA, et al. 2018. Cell. 173:1083. PubMed
  254. Aziz M, et al. 2017. J Immunol. 10.4049/jimmunol.1602056. PubMed
  255. Pascual-Pasto G, et al. 2022. J Immunother Cancer. 10:. PubMed
  256. Lehrke MJ, et al. 2021. Elife. 10:. PubMed
  257. Wang Y, et al. 2021. EMBO Mol Med. 13:e14089. PubMed
  258. Chiang M, et al. 2019. Cell Rep. 28:3212. PubMed
  259. Valdespino-Vázquez MY, et al. 2021. J Med Virol. 93:4480. PubMed
  260. Kissiov DU, et al. 2022. Elife. 11:. PubMed
  261. Isobe T, et al. 2022. Nat Commun. 13:4501. PubMed
  262. Dent JR, et al. 2019. J Appl Physiol (1985). 127:1117. PubMed
  263. Schmidt A, et al. 2017. Toxicol Lett. 10.1016/j.toxlet.2017.08.008. PubMed
  264. Canals Hernaez D, et al. 2022. Front Oncol. 12:856424. PubMed
  265. Mascharak S, et al. 2022. Cell Stem Cell. 29:315. PubMed
  266. Rodriguez A, et al. 2021. Cell Stem Cell. 28:33. PubMed
  267. Pang J, et al. 2021. STAR Protoc. 2:100879. PubMed
  268. Haschak M, et al. 2021. Aging (Albany NY). 13:16938. PubMed
  269. Phan TS, et al. 2021. Sci Adv. 7:. PubMed
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  273. Vergoni B, et al. 2016. Diabetes. 65: 3062 - 3074. PubMed
  274. Burns F, et al. 2015. Aquatic Toxicology. 164: 52-60. PubMed
  275. Stevens M, et al. 2016. Nat Biotechnol. 34:1161-1167. PubMed
  276. Shilts J, et al. 2021. Sci Rep. 0.745138889. PubMed
  277. Jain A, et al. 2020. Nat Immunol. 0.920138889. PubMed
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Antigen Details

Biology Area
Cell Biology, Cell Cycle/DNA Replication, Neuroscience
Antigen References

1. Langlois RG, et al. 1980. Chromosoma. 77:229.
2. Kubista M, et al. 1987. Biochemistry 26:4545.

Gene ID
NA

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Go To Top Version: 2    Revision Date: 04/24/2025

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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Pricing & Availability
Regulatory Status
RUO
Other Names
4',6-Diamidino-2-Phenylindole, Dilactate
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Product Citations
publications
1_Ankita_Patil.jpg
Methanol fixed superior cervical ganglion neurons were stained with anti-tubulin β3 (pink) and anti-TRIM46 (aqua), then stained with secondary antibodies, phalloidin (orange) and DAPI (blue). Image generously submitted to the 2017 Cell Life Imaging Competition by Ankita Patil from Drexel University.
  • 1_Ankita_Patil.jpg
    Methanol fixed superior cervical ganglion neurons were stained with anti-tubulin β3 (pink) and anti-TRIM46 (aqua), then stained with secondary antibodies, phalloidin (orange) and DAPI (blue). Image generously submitted to the 2017 Cell Life Imaging Competition by Ankita Patil from Drexel University.
  • 2_Kathleen-Ignatoski
    Frozen mouse intestine stained with purified anti-Ankyrin-B (green), E-Cadherin (red) and DAPI (blue) followed by anti-goat Alexa Fluor® 488 and anti-rabbit Alexa Fluor® 568. Image generously submitted to the 2017 Cell Life Imaging Competition by Kathleen Ignatoski from University of Michigan.
  • 3_KifLiakath.jpg
    Whole mount mouse tail epidermis was stained with purified anti-Keratin 14 (green, clone Poly19053), purified anti-Keratin 15 (orange, clone Poly18339) and DAPI (blue). Image generously submitted to the 2017 Cell Life Imaging Competition by Kif Liakath.
  • 4_DAPI_IF_080712
    Human cervical cancer cell line, HeLa, was fixed with 4% paraformaldehyde and stained with 300 nM DAPI.
  • 5_DAPI_080712
    Human cervical cancer cell line, HeLa, was fixed with 4% paraformaldehyde and stained with 300 nM DAPI.
See DAPI (4',6-Diamidino-2-Phenylindole, Dilactate) spectral data
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422801 10 mg DKK600
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Description

DAPI (4',6-Diamidino-2-Phenylindole, Dilactate), a blue fluorescent nucleic acid stain, preferentially binds dsDNA and associates with the minor groove AT clusters. The molecular weight for DAPI is 457.5.  Association of DAPI with dsDNA increases fluorescence approximately 20-fold, with an emission maximum of 460 nm. The increase in fluorescence is believed to be due to displacement of water molecules from the minor groove and DAPI. DAPI can also bind RNA and emits at a longer wavelength of 500 nm, with only a 20% increase in fluorescence.

DAPI is a popular nuclear stain for identification of cell cycle and specifically stains nuclei but not cytoplasm. The dilactate form is more water soluble compared to the dihydrochloride form.

DAPI can be excited by a UV laser, xenon, or a mercury-arc lamp. 

Product Details
Technical Data Sheet (pdf)

Product Details

Formulation
Lyophilized
Preparation
To make 5 mg/ml (10.9 mM) solution, dissolve entire contents of vial in 2 ml of deionized water.
Storage & Handling
Store between 2°C and 8°C, protected from light. For long term storage, aliquot and store at -20°C.
Application

ICFC, ICC
Recommended Usage

This product can be used at 30 µM for flow cytometry and 300 nM for immunofluorescence.  It is recommended to titrate the dye for optimal performance. 

Excitation Laser
Ultraviolet Laser (355 nm)
Application Notes

Protocols for Counterstaining with DAPI:

Immunofluorescence Staining
Note: Cells may be fixed by method of choice, such as with 4% paraformaldehyde. DAPI staining is done after staining for other markers. Fixation and permeabilization of cells is not necessary to counterstain with DAPI.

1. Fix cells using method of choice.
2. Incubate the cells with phosphate buffered saline (PBS) for 15 minutes.
3. Dilute the DAPI solution (we recommend to 300 nM) with PBS. Remove excess PBS from the slide and cover with DAPI solution, making sure the cells are completely covered.
4. Incubate for up to 5 minutes. Rinse the slide several times to remove all free DAPI. It is recommended to use a mounting medium with antifade reagent to reduce fluorescence quenching. The slide is ready to view under the microscope with appropriate filters.


Nuclear Staining for Flow Cytometry
Note: Cells may be fixed with 4% paraformadehyde or absolute ethanol.

1. Fix cells with absolute ethanol or 4% paraformaldehyde.
2. Centrifuge, discard the supernatant, and add 5 ml of PBS, allowing cells to rehydrate for 15 minutes.
3. Dilute the DAPI to 30 µM (recommended) in staining buffer (such as BioLegend's FOXP3 Perm Buffer, Cat. No. 421402).
4. Centrifuge cell suspension from step 3 above, discard supernatant, and add 1 ml of DAPI solution; then incubate at room temperature for 15 minutes.
5. Analyze by flow cytometry using an instrument with appropriate lasers.

Additional Product Notes

View more applications data for this product in our Scientific Poster Library.

Application References
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  172. Xiong E, et al. 2023. Front Immunol. 13:1083119. PubMed
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  174. González-Terán B, et al. 2022. Cell. 185:794. PubMed
  175. Wiedemann SJ, et al. 2022. Cell Metab. 34:991. PubMed
  176. Michalek S, et al. 2022. EMBO Rep. 23:e54195. PubMed
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  184. Velilla J, et al. 2019. Neurol Genet. 5:e312. PubMed
  185. Jiao Q, et al. 2017. Clin Exp Immunol. 188:420. PubMed
  186. Rodrigues M, et al. 2022. Biochem J. 479:1891. PubMed
  187. Gabriely G, et al. 2021. iScience. 24:103347. PubMed
  188. Effern M, et al. 2022. STAR Protoc. 3:101038. PubMed
  189. Sountoulidis A, et al. 2020. PLoS Biol. 18:e3000675. PubMed
  190. Kobayashi T, et al. 2019. Cell. 176:982. PubMed
  191. Wilson K, et al. 2016. Cell Death Dis. 7: e2197. PubMed
  192. Anguiano T, et al. 2020. Toxicol Sci. 177:494. PubMed
  193. Afshar Y, et al. 2021. Nat Commun. 12:4408. PubMed
  194. Goodnough LH, et al. 2022. Front Endocrinol (Lausanne). 13:924927. PubMed
  195. Nguyen DT, et al. 2022. Cells. 11:. PubMed
  196. Baldominos P, et al. 2022. STAR Protoc. 3:101795. PubMed
  197. Elias A, et al. 2016. Sci Rep. 6: 24971. PubMed
  198. Miedema A, et al. 2022. Acta Neuropathol Commun. 10:8. PubMed
  199. Emmons TR, et al. 2021. Cancer Immunol Res. 9:790. PubMed
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  201. Gamboa CM, et al. 2021. STAR Protoc. 100345:2. PubMed
  202. Sugawara E, et al. 2020. Autophagy. 1.323611111. PubMed
  203. Brunner JS, et al. 2020. Nat Commun. 0.757638889. PubMed
  204. Fukushima T, et al. 2019. Cell Rep. 29:4144. PubMed
  205. Bannard O, et al. 2016. J Exp Med. 13: 993 - 1009. PubMed
  206. Sumitomo Y, et al. 2016. Blood. 128: 1614 - 1624. PubMed
  207. Paris AJ, et al. 2020. Nat Cell Biol. 1.747916667. PubMed
  208. Zhang M, et al. 2017. Oncol Lett. 3.069444444. PubMed
  209. Greenblatt SM, et al. 2018. Cancer Cell. 33:1111. PubMed
  210. Alexander MS, et al. 2021. Muscle Nerve. 63:928. PubMed
  211. Hendricks A, et al. 2021. Front Oncol. 11:646885. PubMed
  212. Hiramatsu-Asano S, et al. 2021. Front Immunol. 11:616141. PubMed
  213. Ambrosi TH, et al. 2021. Elife. 10: . PubMed
  214. Hopp AK, et al. 2021. Mol Cell. 81:340. PubMed
  215. Allam O, et al. 2018. PLoS One. 13:e0194185. PubMed
  216. DeDreu J, et al. 2020. FASEB J. 34:9316. PubMed
  217. Wilson AS, et al. 2022. Nat Commun. 13:528. PubMed
  218. Deng J, et al. 2021. Breast Cancer Res Treat. 185:85. PubMed
  219. Sodhi C, et al. 2015. J Immunol. 194:4931. PubMed
  220. Kanda K, et al. 2018. JCI Insight. 3:e91316. PubMed
  221. Fujiwara H, et al. 2021. Nat Immunol. 22:1440. PubMed
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Antigen Details

Biology Area
Cell Biology, Cell Cycle/DNA Replication, Neuroscience
Antigen References

1. Langlois RG, et al. 1980. Chromosoma. 77:229.
2. Kubista M, et al. 1987. Biochemistry 26:4545.

Gene ID
NA
Go To Top Version: 2    Revision Date: 04/24/2025

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