Nile Red *CAS 7385-67-3*

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Chemical structure for Nile Red *CAS 7385-67-3*
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25 mg 22190 $75

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Ex/Em (nm)552/636
CAS #7385-67-3
Storage F/D/L
Category Cell Biology
Labeling Cells
Related Lipoproteins
Vital Stains
Nile red (also known as Nile blue oxazone) is a lipophilic stain. It has environment-sensitive fluorescence. Nile red is intensely fluorescent in a lipid-rich environment while it has minimal fluorescence in aqueous media. It is an excellent vital stain for the detection of intracellular lipid droplets by fluorescence microscopy and flow cytofluorometry. Nile red stains intracellular lipid droplets red. Better selectivity for cytoplasmic lipid droplets can be obtained when the cells are viewed for yellow-gold fluorescence (450-500 nm excitation; >528 nm emission) rather than red fluorescence (515-560 nm excitation; >590 nm emission). It is strongly fluorescent, but only in a hydrophobic environment.

Common stock solution preparation

Table 1. Volume of DMSO needed to reconstitute specific mass of Nile Red *CAS 7385-67-3* to given concentration. Note that volume is only for preparing stock solution. Refer to sample experimental protocol for appropriate experimental/physiological buffers.

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Table 2. Enter any two values (mass, volume, concentration) to calculate the third.

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This protocol only provides a guideline, and should be modified according to your specific needs.

1. Prepare Nile Red working solution:

1.1    Prepare a 1 mM stock solution of Nile Red in high-quality, anhydrous DMSO. The stock solution should be used promptly; any remaining solution should be aliquoted and frozen at < -20 oC.

Note: Avoid repeated freeze-thaw cycles, and protect from light.

1.2    Prepare a 200 to 1000 nM working solution of 1X Nile Red right before use by diluting Nile Red DMSO stock solution (from Step 1.1) in Hanks and 20 mM Hepes buffer (HHBS) or buffer of your choice, pH 7. Mix them well by votexing.


2. Run Nile Red with a fluorescence microscope or a flow cytometer:

2.1    Treat cells with test compounds for a desired period of time.

2.2     Centrifuge the cells to get 1-5 x 105 cells per tube.

2.3     Resuspend cells in 500 µL of Nile Red working solution (from Step 1.2).

2.4     Incubate at room temperature or 37° C for 5 to 10 min, protected from light.

2.5     To remove the Nile Red working solution from the cells, wash the cells with HHBS or buffer of your choice. Resuspend cells in 500 µL of prewarmed HHBS or medium to get 1-5 x 105 cells per tube.

2.6     Monitor the fluorescence change at Ex/Em = 552/636 nm with a fluorescence microscope or a flow cytometer.


Note 1: For adherent cells, cells can be washed with HHBS or buffer of your choice, and loaded with Nile Red working solution directly in the cell plate.

Note 2: Cells can also be prefixed, and then stained with Nile Red working solution.

References & Citations

Depot-specific characteristics of adipose tissue-derived stromal cells in thyroid-associated orbitopathy
Authors: Janice Siu Chong Wong, Wai Kit Chu, Benjamin Fuk-Loi Li, Chi-Pui Pang, Kelvin Kam-lung Chong
Journal: British Journal of Ophthalmology (2018): bjophthalmol--2017

Indole-3-acetic-acid-induced phenotypic plasticity in Desmodesmus algae
Authors: Tan-Ya Chung, Chih-Yen Kuo, Wei-Jiun Lin, Wei-Lung Wang, Jui-Yu Chou
Journal: Scientific reports (2018): 10270

CRISPR/CAS9-mediated Tspo gene mutations lead to reduced mitochondrial membrane potential and STEROID FORMation in MA-10 mouse tumor Leydig cells
Authors: Jinjiang Fan, Kevin Wang, Barry Zirkin, Vassilios Papadopoulos
Journal: Endocrinology (2017)

Long-term exposure to abnormal glucose levels alters drug metabolism pathways and insulin sensitivity in primary human hepatocytes
Authors: Matthew D Davidson, Kimberly R Ballinger, Salman R Khetani
Journal: Scientific Reports (2016)

Hormone and drug-mediated modulation of glucose metabolism in a microscale model of the human liver
Authors: Matthew D Davidson, Michael Lehrer, Salman R Khetani
Journal: Tissue Engineering Part C: Methods (2015): 716--725

Hepatocyte nuclear factor 4α and downstream secreted phospholipase A2 GXIIB regulate production of infectious hepatitis C virus
Authors: Xinlei Li, Hanfang Jiang, Linbing Qu, Wenxia Yao, Hua Cai, Ling Chen, Tao Peng
Journal: Journal of virology (2014): 612--627

Characterization of the squalene-rich Botryococcus braunii Abt02 strain
Authors: Min Cao, Fangfang Zhang, Yunxiang Mao, Fanna Kong, Dongmei Wang
Journal: Journal of Oceanology and Limnology: 1--10

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