Nile Red | CAS#7385-67-3 | Fluorescent probe

MedKoo Cat#: 462794 | Name: Nile Red

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Description:

WARNING: This product is for research use only, not for human or veterinary use.

Nile red is a fluorescent probe commonly used for the detection of intracellular lipid droplets. It displays excitation/emission maxima of 529/576 nm, respectively, for neutral lipids. It is selective for neutral lipids at emission wavelengths of 580 nm or lower, but, at higher emission wavelengths, phospholipid membranes are also visualized. Nile red has been used in live, fixed, or unfixed cells with detection by fluorescence microscopy, spectroscopy, or flow cytometry. It has been used as a marker of lipid droplets in cellular models of hepatocyte steatosis.

Chemical Structure

Nile Red

CAS#7385-67-3

Theoretical Analysis

MedKoo Cat#: 462794

Name: Nile Red

CAS#: 7385-67-3

Chemical Formula: C20H18N2O2

Exact Mass: 318.1368

Molecular Weight: 318.38

Elemental Analysis: C, 75.45; H, 5.70; N, 8.80; O, 10.05

Price and Availability

Size	Price	Availability
250mg	USD 350.00	2 Weeks
500mg	USD 550.00	2 Weeks
1g	USD 950.00	2 Weeks

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Synonym

Nile Red;

IUPAC/Chemical Name

9-(diethylamino)-5H-benzo[a]phenoxazin-5-one

InChi Key

VOFUROIFQGPCGE-UHFFFAOYSA-N

InChi Code

InChI=1S/C20H18N2O2/c1-3-22(4-2)13-9-10-16-18(11-13)24-19-12-17(23)14-7-5-6-8-15(14)20(19)21-16/h5-12H,3-4H2,1-2H3

SMILES Code

O=C1C=C2OC3=CC(N(CC)CC)=CC=C3N=C2C4=CC=CC=C14

Appearance

Solid powder

Purity

>98% (or refer to the Certificate of Analysis)

Shipping Condition

Shipped under ambient temperature as non-hazardous chemical. This product is stable enough for a few weeks during ordinary shipping and time spent in Customs.

Storage Condition

Dry, dark and at 0 - 4 C for short term (days to weeks) or -20 C for long term (months to years).

Solubility

Soluble in DMSO

Shelf Life

>3 years if stored properly

Drug Formulation

This drug may be formulated in DMSO

Stock Solution Storage

0 - 4 C for short term (days to weeks), or -20 C for long term (months).

HS Tariff Code

2934.99.9001

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Product Data

Product Data

Instruction

View handling instruction

Biological target:

Nile red is a fluorescent probe commonly used for the detection of intracellular lipid droplets.

In vitro activity:

Nile red-stained, lipid droplet-filled macrophages exhibited greater fluorescence intensity than did nile red-stained control macrophages, and the two cell populations could be differentiated and analyzed by flow cytofluorometry. Nile red exhibits properties of a near-ideal lysochrome. It is strongly fluorescent, but only in the presence of a hydrophobic environment. Reference: J Cell Biol. 1985 Mar;100(3):965-73. https://pubmed.ncbi.nlm.nih.gov/3972906/

In vivo activity:

The labeled and non-labeled micelle formulations were tested in vitro in cells (Rhodamine-micelle formulations), then in vivo in a case study of an ophthalmic application (Nile Red micelle formulations). Both in vitro and in vivo experiments revealed a significant improvement of fluorescence stability of the MPEG-PHOA-dye formulations, facilitating the investigations on tracing the micelles and their stability. Reference: Eur J Pharm Biopharm. 2019 Sep;142:480-487. https://pubmed.ncbi.nlm.nih.gov/31336183/

	Solvent	mg/mL	mM
Solubility
	DMF	1.0	3.14
	DMSO	23.6	73.95
	DMSO:PBS (pH 7.2) (1:10)	0.1	0.28
	Ethanol	1.0	3.14

Note: There can be variations in solubility for the same chemical from different vendors or different batches from the same vendor. The following factors can affect the solubility of the same chemical: solvent used for crystallization, residual solvent content, polymorphism, salt versus free form, degree of hydration, solvent temperature. Please use the solubility data as a reference only. Warming and sonication will facilitate dissolving. Still have questions? Please contact our Technical Support scientists.

Preparing Stock Solutions

The following data is based on the product molecular weight 318.38 Batch specific molecular weights may vary from batch to batch due to the degree of hydration, which will affect the solvent volumes required to prepare stock solutions.

Concentration / Solvent Volume / Mass	1 mg	5 mg	10 mg
1 mM	1.15 mL	5.76 mL	11.51 mL
5 mM	0.23 mL	1.15 mL	2.3 mL
10 mM	0.12 mL	0.58 mL	1.15 mL
50 mM	0.02 mL	0.12 mL	0.23 mL

Formulation protocol:

1. Pino EC, Webster CM, Carr CE, Soukas AA. Biochemical and high throughput microscopic assessment of fat mass in Caenorhabditis elegans. J Vis Exp. 2013 Mar 30;(73):50180. doi: 10.3791/50180. PMID: 23568026; PMCID: PMC3944676. 2. Greenspan P, Mayer EP, Fowler SD. Nile red: a selective fluorescent stain for intracellular lipid droplets. J Cell Biol. 1985 Mar;100(3):965-73. doi: 10.1083/jcb.100.3.965. PMID: 3972906; PMCID: PMC2113505. 3. Bisogno S, Gąsior Ł, Ptak GE. Nile Red and BODIPY Staining of Lipid Droplets in Mouse Oocytes and Embryos. Methods Mol Biol. 2023;2566:205-212. doi: 10.1007/978-1-0716-2675-7_16. PMID: 36152253. 4. Trubitsyn G, Nguyen VN, Di Tommaso C, Borchard G, Gurny R, Möller M. Impact of covalently Nile Red and covalently Rhodamine labeled fluorescent polymer micelles for the improved imaging of the respective drug delivery system. Eur J Pharm Biopharm. 2019 Sep;142:480-487. doi: 10.1016/j.ejpb.2019.07.020. Epub 2019 Jul 20. PMID: 31336183.

In vitro protocol:

In vivo protocol:

1. Bisogno S, Gąsior Ł, Ptak GE. Nile Red and BODIPY Staining of Lipid Droplets in Mouse Oocytes and Embryos. Methods Mol Biol. 2023;2566:205-212. doi: 10.1007/978-1-0716-2675-7_16. PMID: 36152253. 2. Trubitsyn G, Nguyen VN, Di Tommaso C, Borchard G, Gurny R, Möller M. Impact of covalently Nile Red and covalently Rhodamine labeled fluorescent polymer micelles for the improved imaging of the respective drug delivery system. Eur J Pharm Biopharm. 2019 Sep;142:480-487. doi: 10.1016/j.ejpb.2019.07.020. Epub 2019 Jul 20. PMID: 31336183.

1: Wang W, Xing J, Ge Z. Evaluation of Nile Red-Loaded Mesoporous Silica Nanoparticles for Developing Water-Soaked Fingerprints on Thermal Paper. J Forensic Sci. 2019 May;64(3):717-727. doi: 10.1111/1556-4029.13959. Epub 2018 Nov 16. PMID: 30444941. 2: Martinez V, Henary M. Nile Red and Nile Blue: Applications and Syntheses of Structural Analogues. Chemistry. 2016 Sep 19;22(39):13764-13782. doi: 10.1002/chem.201601570. Epub 2016 Jul 13. PMID: 27406265. 3: Korber J, Barth C, Gibbs S. Nile Red derivatives enable improved ratiometric imaging for nerve-specific contrast. J Biomed Opt. 2018 Jul;23(7):1-13. doi: 10.1117/1.JBO.23.7.076002. PMID: 29981230. 4: Trubitsyn G, Nguyen VN, Di Tommaso C, Borchard G, Gurny R, Möller M. Impact of covalently Nile Red and covalently Rhodamine labeled fluorescent polymer micelles for the improved imaging of the respective drug delivery system. Eur J Pharm Biopharm. 2019 Sep;142:480-487. doi: 10.1016/j.ejpb.2019.07.020. Epub 2019 Jul 20. PMID: 31336183. 5: Gagné F, Auclair J, Quinn B. Detection of polystyrene nanoplastics in biological samples based on the solvatochromic properties of Nile red: application in Hydra attenuata exposed to nanoplastics. Environ Sci Pollut Res Int. 2019 Nov;26(32):33524-33531. doi: 10.1007/s11356-019-06501-3. Epub 2019 Oct 2. PMID: 31578681. 6: Zhao C, Luo MT, Huang C, Chen XF, Xiong L, Li HL, Chen XD. Determining intracellular lipid content of different oleaginous yeasts by one simple and accurate Nile Red fluorescent method. Prep Biochem Biotechnol. 2019;49(6):597-605. doi: 10.1080/10826068.2019.1587624. Epub 2019 Mar 31. PMID: 30929602. 7: Alemán-Nava GS, Cuellar-Bermudez SP, Cuaresma M, Bosma R, Muylaert K, Ritmann BE, Parra R. How to use Nile Red, a selective fluorescent stain for microalgal neutral lipids. J Microbiol Methods. 2016 Sep;128:74-79. doi: 10.1016/j.mimet.2016.07.011. Epub 2016 Jul 16. PMID: 27432343. 8: Liu XD, Fan C, Sun R, Xu YJ, Ge JF. Nile-red and Nile-blue-based near- infrared fluorescent probes for in-cellulo imaging of hydrogen sulfide. Anal Bioanal Chem. 2014 Nov;406(28):7059-70. doi: 10.1007/s00216-014-8131-y. Epub 2014 Sep 26. PMID: 25258280. 9: Lan JS, Zeng RF, Liu Y, Xiang YW, Jiang XY, Liu L, Xie SS, Ding Y, Zhang T. A near-infrared Nile red fluorescent probe for the discrimination of biothiols by dual-channel response and its bioimaging applications in living cells and animals. Analyst. 2019 Jun 7;144(11):3676-3684. doi: 10.1039/c9an00280d. Epub 2019 May 14. PMID: 31086902. 10: Balduyck L, Veryser C, Goiris K, Bruneel C, Muylaert K, Foubert I. Optimization of a Nile Red method for rapid lipid determination in autotrophic, marine microalgae is species dependent. J Microbiol Methods. 2015 Nov;118:152-8. doi: 10.1016/j.mimet.2015.09.009. Epub 2015 Sep 24. PMID: 26388510. 11: Kreder R, Pyrshev KA, Darwich Z, Kucherak OA, Mély Y, Klymchenko AS. Solvatochromic Nile Red probes with FRET quencher reveal lipid order heterogeneity in living and apoptotic cells. ACS Chem Biol. 2015 Jun 19;10(6):1435-42. doi: 10.1021/cb500922m. Epub 2015 Mar 6. PMID: 25710589. 12: Nath A, Fernández C, Lampe JN, Atkins WM. Spectral resolution of a second binding site for Nile Red on cytochrome P4503A4. Arch Biochem Biophys. 2008 Jun 1;474(1):198-204. doi: 10.1016/j.abb.2008.03.017. Epub 2008 Mar 25. PMID: 18395506; PMCID: PMC2474736. 13: Prioli S, Reinholdt P, Hornum M, Kongsted J. Rational Design of Nile Red Analogs for Sensing in Membranes. J Phys Chem B. 2019 Dec 12;123(49):10424-10432. doi: 10.1021/acs.jpcb.9b09691. Epub 2019 Dec 4. PMID: 31730348. 14: Bartel M, Wysocka B, Krug P, Kępińska D, Kijewska K, Blanchard GJ, Kaczyńska K, Lubelska K, Wiktorska K, Głowala P, Wilczek M, Pisarek M, Szczytko J, Twardowski A, Mazur M. Magnetic polymer microcapsules loaded with Nile Red fluorescent dye. Spectrochim Acta A Mol Biomol Spectrosc. 2018 Apr 15;195:148-156. doi: 10.1016/j.saa.2018.01.056. Epub 2018 Jan 31. PMID: 29414572. 15: Lampe JN, Fernandez C, Nath A, Atkins WM. Nile Red is a fluorescent allosteric substrate of cytochrome P450 3A4. Biochemistry. 2008 Jan 15;47(2):509-16. doi: 10.1021/bi7013807. Epub 2007 Dec 20. PMID: 18092806. 16: Liu K, Kong X, Ma Y, Lin W. Preparation of a Nile Red-Pd-based fluorescent CO probe and its imaging applications in vitro and in vivo. Nat Protoc. 2018 May;13(5):1020-1033. doi: 10.1038/nprot.2018.013. Epub 2018 Apr 19. PMID: 29674754. 17: Escorcia W, Ruter DL, Nhan J, Curran SP. Quantification of Lipid Abundance and Evaluation of Lipid Distribution in Caenorhabditis elegans by Nile Red and Oil Red O Staining. J Vis Exp. 2018 Mar 5;(133):57352. doi: 10.3791/57352. PMID: 29553519; PMCID: PMC5931440. 18: Diaz G, Melis M, Batetta B, Angius F, Falchi AM. Hydrophobic characterization of intracellular lipids in situ by Nile Red red/yellow emission ratio. Micron. 2008 Oct;39(7):819-24. doi: 10.1016/j.micron.2008.01.001. Epub 2008 Jan 11. PMID: 18329888. 19: Gattuso H, Besancenot V, Grandemange S, Marazzi M, Monari A. From non- covalent binding to irreversible DNA lesions: nile blue and nile red as photosensitizing agents. Sci Rep. 2016 Jun 22;6:28480. doi: 10.1038/srep28480. PMID: 27329409; PMCID: PMC4916457. 20: Uehara N, Yoshida O. Release of Nile red from thermoresponsive gold nanocomposites by heating a solution and the addition of glutathione. Anal Sci. 2012;28(12):1125-32. doi: 10.2116/analsci.28.1125. PMID: 23232230.