Reagents and instruments for immunology, cell biology and molecular biology.

Nile Red [7385-67-3]

Print

Cat# HY-D0718-100mg

Size : 100mg

Brand : MedChemExpress

Request more information

Description

Nile red (Nile blue oxazone) is a lipophilic stain. Nile red has environment-sensitive fluorescence. Nile red is intensely fluorescent in a lipid-rich environment while it has minimal fluorescence in aqueous media. Nile red is an excellent vital stain for the detection of intracellular lipid droplets by fluorescence microscopy and flow cytof uorometry. Nile red stains intracellular lipid droplets red. The fluorescence wavelength is 559/635 nm^[1].

In Vitro

1.1 Preparation of stock solution
Use DMSO to prepare 1 mM stock solution.
1.2 Preparation of working solution
Use preheated serum-free cell culture medium or PBS to dilute the stock solution to a final concentration of 200-1000 nM.
Note: Please adjust the concentration of Nile Red working solution according to the actual situation and prepare it before use.
2. Cell staining
2.1 Suspended cells: Collect cells by centrifugation and wash twice with PBS for 5 minutes each time.
Adherent cells: Discard the culture medium and add trypsin to digest the cells. After centrifugation and discarding the supernatant, wash twice with PBS for 5 minutes each time.
2.2 Add 1 mL Nile Red working solution and incubate at room temperature for 5-10 minutes.
2.3 Centrifuge at 400 g, 4℃ for 3-4 minutes and discard the supernatant.
2.4 Wash cells twice with PBS for 5 minutes each time.
2.5 Resuspend the cells in 1 mL serum-free medium or PBS and observe under a fluorescence microscope.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

In Vivo

When Nile red-stained Caenorhabditis elegans is viewed for green fluorescence, discrete lipid bodies can be observed throughout the intestine and other tissues either in clusters or evenly dispersed, depending on the animal's genotype or experimental treatment^[3].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Molecular Weight

318.37

Formula

C₂₀H₁₈N₂O₂

CAS No.

7385-67-3

Appearance

Solid

Color

Green to dark green

Emission (Em)

598

Excitation (Ex)

543

SMILES

O=C1C2=CC=CC=C2C3=NC4=CC=C(N(CC)CC)C=C4OC3=C1

Shipping

Room temperature in continental US; may vary elsewhere.

Storage

4°C, protect from light

*In solvent : -80°C, 2 years; -20°C, 1 year (protect from light)

Solvent & Solubility

In Vitro:

DMSO : 2 mg/mL (6.28 mM; ultrasonic and warming and heat to 60°C; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

Ethanol : 1 mg/mL (3.14 mM; ultrasonic and warming and heat to 60°C)

Preparing
Stock Solutions

Concentration Solvent Mass	1 mg	5 mg	10 mg

1 mM	3.1410 mL	15.7050 mL	31.4100 mL
5 mM	0.6282 mL	3.1410 mL	6.2820 mL
10 mM	---	---	---

View the Complete Stock Solution Preparation Table

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 2 years; -20°C, 1 year (protect from light). When stored at -80°C, please use it within 2 years. When stored at -20°C, please use it within 1 year.

Select the appropriate dissolution method based on your experimental animal and administration route.

For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

Protocol 1

Add each solvent one by one: 10% DMSO 40% PEG300 5% Tween-80 45% Saline
Solubility: 0.2 mg/mL (0.63 mM); Suspended solution; Need ultrasonic

This protocol yields a suspended solution of 0.2 mg/mL. Suspended solution can be used for oral and intraperitoneal injection.
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (2.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.
Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
Protocol 2

Add each solvent one by one: 10% DMSO 90% (20% SBE-β-CD in Saline)
Solubility: ≥ 0.2 mg/mL (0.63 mM); Clear solution

This protocol yields a clear solution of ≥ 0.2 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (2.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.