CAS 7385-67-3 Nile Red - Analytical Products / Alfa Chemistry

CAT	AP7385673
CAS	7385-67-3
Structure
MDL Number	MFCD00011639
Molecular Weight	318.37
EC Number	230-966-0
InChI Key	VOFUROIFQGPCGE-UHFFFAOYSA-N
REAXYS Number	279110

How to Use Nile Red, a Selective Fluorescent Stain for Microalgal Neutral Lipids

Gibrán S Alemán-Nava, Sara P Cuellar-Bermudez, María Cuaresma, Rouke Bosma, Koenraad Muylaert, Bruce E Ritmann, Roberto Parra

J Microbiol Methods. 2016 Sep;128:74-79.

PMID: 27432343

Nile Red and Nile Blue: Applications and Syntheses of Structural Analogues

Vincent Martinez, Maged Henary

Chemistry. 2016 Sep 19;22(39):13764-13782.

PMID: 27406265

Nile Red Derivatives Enable Improved Ratiometric Imaging for Nerve-Specific Contrast

Jesse Korber, Connor Barth, Summer Gibbs

J Biomed Opt. 2018 Jul;23(7):1-13.

PMID: 29981230

Nile Red: A Selective Fluorescent Stain for Intracellular Lipid Droplets

P Greenspan, E P Mayer, S D Fowler

J Cell Biol. 1985 Mar;100(3):965-73.

PMID: 3972906

Quantification of Lipid Abundance and Evaluation of Lipid Distribution in Caenorhabditis Elegans by Nile Red and Oil Red O Staining

Wilber Escorcia, Dana L Ruter, James Nhan, Sean P Curran

J Vis Exp. 2018 Mar 5;(133):57352.

PMID: 29553519

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CAT	Size	Form	Description	Price
AP7385673-1	10MG		suitable for fluorescence, BioReagent, ≥98.0% (HPLC)	Inquiry
AP7385673-2	100MG, 1G	powder	Technical grade	Inquiry
AP7385673-3	100MG, 1G		for microscopy	Inquiry

Case Study

Nile Red Is Used for the Preparation of Environment-Sensitive GPCR Ligands for No-Wash Fluorescence Imaging

Hanser, Fabien, et al. ACS Chemical Biology 16.4 (2021): 651-660.

Nile Red, a solvatochromic environment-sensitive dye, has been successfully employed for the development of fluorescent probes targeting the oxytocin receptor (OTR), a G protein-coupled receptor (GPCR), under no-wash imaging conditions. In this study, Nile Red was conjugated to the high-affinity nonpeptidic OTR antagonist PF-3274167 via polyethylene glycol (PEG) linkers of varying lengths (PEG4-PEG20), yielding a series of OTR-specific fluorescent ligands (Pf-PEG4-NR, Pf-PEG8-NR, Pf-PEG12-NR, Pf-PEG20-NR).
Key to the synthesis was the preparation of Nile Red-based carboxylic acid (NR-COOH), achieved by optimizing the oxidative cyclization of crude nitrosophenol with 1,6-dihydroxynaphthalene, bypassing the problematic recrystallization step to improve yields and reproducibility. This phenol derivative was then alkylated and deprotected to afford NR-COOH in 84% overall yield.
Coupling of NR-COOH to amine-terminated PEGylated derivatives of PF-3274167 was facilitated via standard peptide coupling techniques. The resulting conjugates localized within lipid bilayers near the OTR, enabling ratiometric fluorescence microscopy to probe receptor microenvironments such as local polarity and lipid order.
This application highlights Nile Red's critical role as a fluorophore in designing functional molecular tools for live-cell imaging, particularly in studying endogenous GPCRs within native membranes, offering a powerful platform for advancing receptor pharmacology and membrane biology.

Nile Red Used for the Construction of H₂Sn-Responsive Fluorescent Probe NR-BS4 for In Vivo Imaging

Jun-Ying, L. V., et al. Chinese Journal of Analytical Chemistry 50.9 (2022): 100140.

Nile Red (NR), a hydrophobic fluorophore with strong environmental sensitivity, was employed as the fluorophore scaffold for synthesizing a series of H₂Sn-responsive fluorescent probes, termed NR-BS. This study focused on engineering probes capable of selectively detecting hydrogen polysulfides (H₂Sn, n > 1), critical signaling molecules involved in cellular redox processes.
To achieve this, 2- or 3-hydroxy-substituted Nile Red derivatives were coupled with 14 benzenesulfonyl analogs to form sulfonate esters, exploiting H₂Sn's nucleophilic reactivity. The substituent type and position on the benzene ring significantly influenced the probe's performance. After systematic photophysical characterization, NR-BS4-constructed using 3-hydroxy Nile Red and 2-nitrobenzenesulfonyl-emerged as the optimal candidate.
R-BS4 demonstrated excellent analytical features, including a wide linear detection range (0-350 μM), high sensitivity (LOD = 0.140 μM), and broad pH tolerance (pH 4-10). Importantly, it exhibited minimal interference from biothiols. Density functional theory (DFT) calculations and LC-MS confirmed its PET-based sensing mechanism. Cytotoxicity assays validated the probe's biocompatibility, and fluorescence imaging confirmed its capability for real-time detection of endogenous and exogenous H₂Sn in living cells and zebrafish.
This work highlights Nile Red's versatility as a scaffold for developing selective and sensitive probes, offering valuable tools for studying H₂Sn biology and related redox signaling in complex biological environments.

Nile Red Used for the Preparation of Turn-On Fluorescent Carbon Dots for α-Synuclein Fibril Detection

Li, Qin-Ying, et al. Sensors and Actuators B: Chemical 412 (2024): 135785.

In this study, Nile Red (NR) was employed as a fluorescent carbon source to prepare NR-based carbon dots (NR-CDs) via a one-pot solvothermal synthesis for the selective detection of α-synuclein fibrils. A solution of citric acid (0.21 g in 1 mL deionized water) was mixed with 4 mL ethanol containing 1 mM NR. The mixture was sealed in a 25 mL Teflon-lined autoclave and heated at 180 °C. After the reaction, the product was purified through dialysis (500 Da, 12 h) to remove unreacted precursors.
The NR-CDs exhibited microenvironment-sensitive fluorescence, with red fluorescence significantly enhanced in response to increased viscosity and decreased polarity-characteristics aligned with the fibrillar form of α-synuclein. Notably, the NR-CDs did not respond to the monomeric form, enabling selective turn-on fluorescence detection and imaging of α-synuclein fibrils both in vitro and in vivo. Further optimization studies varying citric acid content and reaction time confirmed the ideal synthesis parameters, resulting in NR-CDs with excitation/emission maxima at 545/630 nm.
Compared to free NR dye, NR-CDs showed superior water dispersibility, lower cytotoxicity, and enhanced photostability. Additionally, NR-CDs demonstrated inhibitory effects on α-synuclein aggregation, indicating potential therapeutic applications. This approach highlights the utility of Nile Red in constructing biocompatible, amyloid-specific fluorescent probes for biomedical imaging and neurodegenerative disease research.