Nile Red
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| Catalog No.GC15539 |
El rojo del Nilo es un tinte común para lípidos.
Products are for research use only. Not for human use. We do not sell to patients.
Cas No.: 7385-67-3
Sample solution is provided at 25 µL, 10mM.
Nile red is a common lipid dye. Nile red is a hydrophobic and metachromatic dye with poor solubility and fluorescence in water, with colour emission varying from deep red to strong yellow gold in hydrophobic environments [1].
Nile red can dye the cholesterol in the human plasma through staining of lipid vesicles in smooth muscle cells and in cultured macrophages incubated at low density, using the excitation/emission wavelengths 450 to 500/>528 [2]. Nile red was also used to study membrane heterogeneity [3] and ligand-hydrophobic protein surface interactions with the alternative wavelengths 570/610 [4] and to study enzyme mechanism by using the wavelengths 550/640 to 660 [5]. Nile red has also been successfully used to stain intracellular neutral lipids that is, TAG and cholesterol esters in yeast, fungi with coupled wavelengths 488/565 to 585 [6] and also in microalgae, with wavelengths set to 488 to 525/570 to 600 [7] or to stain total lipids with wavelengths set to 490/585 [8].
References:
[1]. Rumin J, Bonnefond H, Saint-Jean B, et al. The use of fluorescent Nile red and BODIPY for lipid measurement in microalgae[J]. Biotechnology for biofuels, 2015, 8(1): 1-16.
[2]. Greenspan P, Mayer E P, Fowler S D. Nile red: a selective fluorescent stain for intracellular lipid droplets[J]. The Journal of cell biology, 1985, 100(3): 965-973.
[3]. Ira and, Krishnamoorthy G. Probing the link between proton transport and water content in lipid membranes[J]. The Journal of Physical Chemistry B, 2001, 105(7): 1484-1488.
[4]. Sackett D L, Wolff J. Nile red as a polarity-sensitive fluorescent probe of hydrophobic protein surfaces[J]. Analytical biochemistry, 1987, 167(2): 228-234.
[5]. Ruvinov S B, Yang X J, Parris K D, et al. Ligand-mediated Changes in the Tryptophan Synthase Indole Tunnel Probed by Nile Red Fluorescence with Wild Type, Mutant, and Chemically Modified Enzymes (∗)[J]. Journal of Biological Chemistry, 1995, 270(11): 6357-6369.
[6]. Kimura K, Yamaoka M, Kamisaka Y. Rapid estimation of lipids in oleaginous fungi and yeasts using Nile red fluorescence[J]. Journal of microbiological methods, 2004, 56(3): 331-338.
[7]. Cooksey K E, Guckert J B, Williams S A, et al. Fluorometric determination of the neutral lipid content of microalgal cells using Nile Red[J]. Journal of microbiological methods, 1987, 6(6): 333-345.
[8]. Lee S J, Yoon B D, Oh H M. Rapid method for the determination of lipid from the green alga Botryococcus braunii[J]. Biotechnology techniques, 1998, 12(7): 553-556.
This plan only provides a guide, please modify it to meet your specific needs.
1. Prepare staining solution
(1) Prepare stock solution: Dissolve Nile Red in DMSO and prepare a stock solution with a concentration of 1mM.
Note: Unused stock solution should be aliquoted and stored in the dark at -20°C or -80°C to avoid repeated freezing and thawing.
(2) Prepare working solution: Dilute the stock solution with a suitable buffer (such as serum-free medium or PBS) and prepare a working solution with a concentration of 0.2-1μM.
Note: Please adjust the concentration of the working fluid according to the actual situation and prepare it now.
2. Cell suspension staining
(1) Suspension cells: Centrifuge at 1000g for 3-5 minutes, discard the supernatant, and wash twice with PBS for 5 minutes each time.
(2) Adherent cells: Wash twice with PBS, add trypsin to digest the cells, and centrifuge at 1000g for 3-5 minutes after digestion is completed.
(3) Add 1 mL of Nile Red working solution to resuspend the cells, and incubate at room temperature in the dark for 5-30 minutes. The optimal incubation time for different cells is different, please explore by yourself according to the specific experimental needs.
(4) After the incubation, centrifuge at 1000g for 5 minutes, remove the supernatant, add PBS and wash 2-3 times, 5 minutes each time.
(5) Resuspend cells in pre-warmed serum-free cell culture medium or PBS and observe by fluorescence microscope or flow cytometry.
3. Cell adhesion staining
(1) Culture adherent cells on sterile coverslips.
(2) Remove the coverslip from the culture medium, suck out the excess culture medium, and place the coverslip in a humid environment.
(3) Add 100 μL of dye working solution from one corner of the coverslip and shake gently to evenly cover all cells with the dye.
(4) Incubate at room temperature in the dark for 5-30 minutes. The optimal incubation time for different cells is different, please explore by yourself according to the specific experimental needs.
(5) After the incubation, discard the dye working solution and use pre-warmed culture solution to wash the coverslip 2 to 3 times.
4. Microscopic examination. In triglycerides (neutral lipids), the maximum excitation light/emission light of Nile Red is 515/585nm; in phospholipids (polar lipids), the maximum excitation light/emission light is 554nm/638 nm.
Precautions:
1) Fluorescent dyes all have quenching problems. Please avoid light as much as possible to slow down fluorescence quenching.
2) For your safety and health, please wear a lab coat and disposable gloves.
| Cas No. | 7385-67-3 | SDF | |
| Chemical Name | 9-(diethylamino)-5H-benzo[a]phenoxazin-5-one | ||
| Canonical SMILES | CCN(C1=CC2=C(N=C3C4=CC=CC=C4C(C=C3O2)=O)C=C1)CC | ||
| Formula | C20H18N2O2 | M.Wt | 318.37 |
| Solubility | DMSO : ≥ 50 mg/mL (157.05 mM) | Storage | Store at 4°C, protect from light |
| General tips | Please select the appropriate solvent to prepare the stock solution according to the
solubility of the product in different solvents; once the solution is prepared, please store it in
separate packages to avoid product failure caused by repeated freezing and thawing.Storage method
and period of the stock solution: When stored at -80°C, please use it within 6 months; when stored
at -20°C, please use it within 1 month. To increase solubility, heat the tube to 37°C and then oscillate in an ultrasonic bath for some time. |
||
| Shipping Condition | Evaluation sample solution: shipped with blue ice. All other sizes available: with RT, or with Blue Ice upon request. | ||
| Prepare stock solution | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 3.141 mL | 15.705 mL | 31.41 mL |
| 5 mM | 0.6282 mL | 3.141 mL | 6.282 mL |
| 10 mM | 0.3141 mL | 1.5705 mL | 3.141 mL |
Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)
g
μL
Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)
Calculation results:
Working concentration: mg/ml;
Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )
Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O, mix and clarify.
Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.
Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.
Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
-
Related Biological Data
24HC induces cholesterol accumulation in multivesicular bodies or late endosomes.C. N2a cells were treated with ETOH, 24HC (5 µM), or 25HC (5 µM) for 24 h, then fixed and stained with Nile red. The cells were then analyzed for PE fluorescence intensity by flow cytometr.
N2a cells were treated with ETOH, 24HC (5 µM), or 25HC (5 µM) for 24 h, then fixed and stained with Nile red(GlpBio).
Redox Biology (2023): 102769. PMID: 37285742 IF: 10.7865 -
Related Biological Data
Effects of OG on lipid synthesis in the damaged epidermal barrier. (A, B) Nile red fluorescence staining and quantitative analysis of neutral lipid in the WT (A) and Dectin-1(B) mice.
The stained slices were then stained with Nile Red Fluorescent Dye Working Solution (Glpbio) for 20 min, and finally mounted with glycerol gelatin and examined with a confocal laser scanning microscope.
Brit J Pharmacol (2023). PMID: 38124222 IF: 7.3003 -
Related Biological Data
Effects of SIP on keratinocyte lipid synthesis. (A) Changes in neutral lipid level of HaCaT cells as examined by Nile red fluorescence staining. The plots beside the images show the quantitation analysis of neutral lipids fluorescence intensity. (B) Detection of lipid content in HaCaT cells by oil-red O.
The slices were incubated with Hoechst solution and then in Nile Red Fluorescent Dye Working Solution (Glpbio).
Frontiers in Pharmacology, 2023, 14. PMID: 36733502 IF: 5.9879 -
Related Biological Data
Oil Red O staining, Nile Red staining, and quantifcation of Oil Red O staining in 3T3-L1 cells treated with Citrus sunki peel extract. (a) 3T3-L1 cells, stained with Oil Red O, were visualized in optical microscopy. (b) 3T3-L1 cells, stained with Nile Red, were visualized in optical microscopy.
Nile Red working solution was prepared by diluting nile red dye (GLPBIO) in PBS to a concentration of 1 μg/mL. Cells were treated with nile red working solution for 30 min at room temperature.
J Food Biochem 2024 (2024). IF: 4 -
Related Biological Data
Downregulation of LIPT1 reduces lipid deposition in HCC cells. A Nile red staining analysis showed a signifcant decrease in lipid content in LIPT1-silenced cells.
Briefy, the transfected cells in 96-well plates (3× 103 cells/well) were fixed in 4% paraformaldehyde for 30 min and incubated with 0.1 μg/mL Nile red(GlpBio) for 10 min, then washing the plates with PBS twice, followed by DAPI staining for 10 min.
J Cancer Res Clin (2023): 1-17. PMID: 37668796 IF: 3.5999
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