Nile Red Staining Solution (100 μg/mL) - BioReagent

Cat. No.: N1528845

GRADE & PURITY BioReagent

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Vue d’ensemble

This product is a ready-to-use Nile Red stock staining solution. Nile Red is a lipophilic fluorescent dye that specifically labels lipid droplets and neutral lipids in cells and tissues, emitting orange-red to red fluorescence. It is widely used in microbial lipid detection, plant oil body observation, animal cell lipid droplet fluorescence imaging, and lipid metabolism-related research. Note: This product is a fluorescent dye and is highly sensitive to light. All operations should be performed under light-protected conditions. The dye tends to adsorb to plastic surfaces; glassware is recommended. For research use only. Not for clinical diagnosis.

Specifications

Spécifications et pureté

BioReagent

Conditions de stockage de stockage

Store at -20°C

Expédié en

Ice chest + Ice pads

Ce produit nécessite l'expédition en chaîne froide. Les services terrestres et autres services économiques ne sont pas disponibles.

Grade

BioReagent

Product Line

Produits biologiques

Application Protocols

Tested/validated application details for this specific catalog item are not provided. Only item-specific protocols should be listed here, and they are Not specified for this item; refer to any protocol sheet enclosed with the product or contact Technical Support.

General literature protocols for Nile Red staining exist, but they are not part of this product’s validated instructions.

Biological Roles

Nile Red is not a native biomolecule but functions as an extrinsic fluorescent probe for hydrophobic domains in biological systems.

General/literature notes on behavior in biological contexts:

• Selective partitioning: strongly accumulates in neutral lipid droplets (triacylglycerols, cholesteryl esters) and, to a lesser extent, in certain membrane regions with low polarity. Fluorescence intensity and emission maxima report on microenvironment polarity.
• Minimal aqueous signal: in cytosol or aqueous buffer, emission is weak due to nonradiative decay; this underlies its high contrast for lipid structures.
• Live vs. fixed cells: low-percentage DMSO or alcoholic carriers deliver dye to live cells with limited toxicity when optimized; fixation/permeabilization can alter lipid distribution and thus Nile Red staining patterns.
• Multiplexing: emission shifts enable spectral separation from green and far-red probes; however, overlap can occur with other orange/red fluorophores. Spectral unmixing may be beneficial in complex samples.
• Quantitative use: With standardized acquisition parameters, fluorescence intensity can serve as a relative proxy for lipid content, subject to calibration and control experiments.

Caveats:

• Detergents and strong solvents can extract or redistribute lipids, altering signal localization.
• Photobleaching and self-quenching may occur at high local dye concentrations; titrate to optimal levels.

All uses are for research purposes only (item-specific Research Use Note).

Buffer Applications

This product is a fluorescent staining solution, not a buffering agent. It is not typically used to prepare or adjust buffer pH.

Practical note:

• When diluting to working concentrations, users commonly add the dye stock to pre-made biological buffers (e.g., PBS, HBSS) at very low percentages of organic carrier to deliver the dye. The specific solvent composition of this item is Not specified; confirm buffer compatibility via CoA/SDS and small-scale tests.

Green Alternatives

Nile Red itself is a specialized fluorescent dye; sustainability considerations primarily relate to the solvent system used for stock/working solutions.

Comparison of common carrier solvents (literature, generalized):

• Ethanol/isopropanol

  • Pros: bio-based availability, lower toxicity than many aprotic solvents, readily biodegradable, easy waste management; low cost.
  • Cons: flammable; higher autofluorescence than DMSO in some setups; can fix/permeabilize cells at higher %.

• DMSO

  • Pros: excellent solubilizing power, miscible with water, low volatility; typically low autofluorescence in visible range.
  • Cons: penetrates skin; potential biological effects at >0.5–1% v/v; derived from petroleum processes.

• Acetone

  • Pros: highly volatile (quick drying for fixed samples), effective lipid solvent.
  • Cons: strong defatting action; higher volatility/flammability; can extract lipids aggressively from biological samples.

Greener practice recommendations:

• Use the minimum effective fraction of organic carrier in aqueous working solutions (often ≤0.5–1% v/v).
• Prefer ethanol or isopropanol where compatible with your biology and imaging needs; they offer better EHS profiles than many aprotic alternatives.
• Scale experiments to minimize dye/solvent usage; collect solvent wastes separately for appropriate recycling/disposal.
• Validate reusable staining protocols and imaging conditions to reduce repeats and material consumption.

Note: The specific solvent for this item is Not specified; consult the SDS to align greener substitutions with product compatibility.

Pharmaceutical Uses

This product is for research use only and is not intended for pharmaceutical formulation or clinical applications.

General context (non-clinical):

• Nile Red is occasionally used in pharmaceutical research workflows (e.g., assessing lipid accumulation in cell models, characterizing lipid-based delivery systems, or studying excipient–lipid interactions) as a fluorescent probe. These are laboratory investigations, not drug product uses.

Regulatory/compendial status:

• No pharmacopeial status is implied. Grade is BioReagent (item-specific), suitable for research and analytical studies, not GMP manufacturing.

If use in regulated environments is contemplated, obtain the lot-specific CoA/Spec Sheet and SDS to assess suitability and define incoming specifications (solvent identity, dye concentration, impurities, stability) for your quality system.

Physical Properties

Item-specific physicochemical specifications (BP/MP/density/solvent matrix) are not provided for this ready-to-use solution. See the CoA/Spec Sheet for definitive values.

General/literature properties for Nile Red dyes and their solutions (for method planning only):

• Solubility (literature): readily soluble in many organic solvents (e.g., DMSO, acetone, ethanol, isopropanol); very low solubility in water. In aqueous samples, staining relies on partitioning of the dye into hydrophobic phases (lipid droplets/membranes).
• Photophysics (solvatochromic; literature ranges):
  • Excitation maximum typically in the green–yellow region; commonly excited with 488–561 nm laser lines (e.g., 514 or 543–552 nm).
  • Emission maximum shifts with microenvironment polarity: ~570–590 nm in nonpolar lipids (neutral LDs), progressively red-shifting to ~610–650+ nm in more polar environments. Emission is weak/quenched in bulk aqueous phase.
• Partition behavior (literature): strong preferential accumulation in neutral lipid droplets and other hydrophobic domains; minimal staining of nucleic acids/proteins in aqueous environments.
• Photostability (literature): moderate; solutions and stained samples are light-sensitive and can photobleach under intense illumination.

Notes for users:

• The solvent system, refractive index, pH (if any), and stabilizers of this specific 100 μg/mL solution are Not specified for this item; refer to CoA/Spec Sheet.
• Handle as a light-sensitive fluorescent reagent; minimize exposure to strong light and elevated temperature.

Quality and Grades

• Grade: BioReagent (item-specific). BioReagent-grade materials are intended for research and laboratory applications, commonly conforming to internal quality attributes such as consistency, low particulate burden, and suitability for biological assays. They are not certified for clinical/diagnostic use.
• Purity: Not specified for this item; refer to CoA/Spec Sheet.
• Stabilizers/Preservatives: Not specified for this item; refer to CoA/Spec Sheet.
• UV/fluorescence background: For fluorescence applications, low background and consistent photophysical performance are critical. BioReagent-grade staining solutions are prepared to support reproducible staining intensity and spectral behavior across batches; however, specific spectral cutoff/absorbance specs are Not specified for this item; refer to CoA/Spec Sheet.

What this means for your work:

• Batch-to-batch consistency of the fluorescent response is prioritized; verify excitation/emission settings with your instrument using a small pilot test for optimal detector gain and filter set.
• If your application demands defined solvent composition, water content, or trace impurity limits (e.g., for mass-spec–coupled imaging), request the CoA/Spec Sheet for this lot.
• Research Use Only (item-specific): not for human or animal therapeutic/diagnostic use.

Related practical notes:

• Ready-to-use concentration (100 μg/mL) reduces handling variability relative to preparing solid dye stocks. If different working concentrations are required, perform serial dilutions using a solvent compatible with the original matrix (solvent identity Not specified for this item; confirm via CoA/SDS).

Reaction and Applications

This product is used as a fluorescent staining reagent rather than as a synthetic reactant.

Key applications (literature/general):

• Neutral lipid and lipid droplet staining in cells, tissues, and microorganisms; robust signal in hydrophobic environments with minimal background in aqueous compartments.
• Flow cytometry and fluorescence microscopy for lipid content quantification and visualization (e.g., adipogenesis studies, metabolic assays, yeast/bacterial lipid bodies).
• Polymer science/materials: probe for microphase separation, hydrophobic domain mapping, and solvent quality via solvatochromic shifts.
• Environmental/colloid science: detection of microplastics and hydrophobic particulates by fluorescence microscopy.

Practical tips (general):

• Light sensitivity: prepare and handle solutions under reduced light; protect samples with amber containers/foil.
• Working concentration: empirical optimization is recommended. Typical final dye levels fall in the low ng/mL to low μg/mL range depending on sample and modality; start with small pilot ranges to avoid over-staining and self-quenching.
• Incubation: allow sufficient time for partitioning into lipid domains (minutes to tens of minutes). Excessive exposure to organic cosolvent should be avoided for live samples.
• Wash steps: brief rinses with buffer can reduce background. Avoid strong detergents that may extract lipids and alter localization.
• Imaging: select excitation in the 488–561 nm range and wide emission collection (e.g., 570–650+ nm) to capture polarity-dependent shifts; adjust detector gain to prevent saturation in bright lipid droplets.

Refer to this product’s CoA/SDS for solvent composition and any specific handling constraints.

Reaction Conditions

This section is interpreted as practical usage conditions for staining (not chemical reactions), since the product is a fluorescent dye solution.

General staining conditions (literature guidance; optimize per system):

• Working concentration: often 10–500 ng/mL for microscopy; up to low μg/mL for flow cytometry or low-signal samples. Start with a small titration around 100 ng/mL to 1 μg/mL.
• Solvent delivery: add from concentrated stock into buffer to achieve ≤0.5–1% v/v organic cosolvent (minimize biological perturbation). The solvent identity for this item is Not specified; consult SDS.
• Incubation: 5–30 minutes at ambient temperature is common for fixed cells/tissues; live cells may require shorter exposure. Protect from light.
• Wash: brief rinses with buffer can lower background while retaining lipid droplet signal.
• Excitation/emission: excite with 488–561 nm lines; collect emission broadly from ~570 to 650+ nm. Adjust filters/gain to capture polarity-dependent shifts without saturation.
• Controls: include solvent-only and known lipid-positive controls to benchmark signal and exclude autofluorescence artifacts.

Expected performance notes:

• Signal increases with lipid content; however, overloading dye can cause self-quenching and increased background. Titrate to find the linear range for your system.

These are general guidelines; definitive, item-specific conditions are not specified and should be established empirically in your laboratory.

Safety and Handling

Authoritative hazard classification (GHS signal word, H-statements, pictograms) is Not specified for this item; refer to the product SDS.

General laboratory safety guidance for Nile Red staining solutions (context, not a substitute for SDS):

• Potential hazards (literature/typical):
  • Many Nile Red stock/working solutions are prepared in flammable organic solvents (e.g., acetone, ethanol, isopropanol, DMSO blends). Treat unknown solvent systems as potentially flammable and harmful if inhaled/ingested or upon skin/eye contact.
  • Dyes can cause staining and may be irritants. Avoid inhalation of aerosols/mists and contact with skin/eyes.
• PPE: lab coat, nitrile gloves, and splash goggles. Use in a chemical fume hood when preparing dilutions or handling bulk volumes.
• Incompatibilities (general): strong oxidizers; strong acids/bases can degrade fluorophores. Avoid exposure to strong light sources to prevent photodegradation.
• First aid (general):
  • Skin: wash with soap and water. Remove contaminated clothing.
  • Eyes: rinse cautiously with water for several minutes; remove contact lenses if present and easy to do; seek medical attention if irritation persists.
  • Inhalation: move to fresh air; seek medical advice if symptoms occur.
  • Ingestion: rinse mouth; do not induce vomiting; seek medical attention.
• Spills: absorb with inert material, ventilate area, dispose as chemical waste per institutional guidelines.
• Waste: Collect dye/organic solvent waste in light-protected, properly labeled containers for hazardous waste disposal.

Always consult the SDS for definitive, product-specific hazard and response information.

Solvent Selection

This product is supplied as a ready-to-use staining solution (100 μg/mL). The exact solvent system is Not specified for this item; refer to CoA/SDS.

General solvent considerations for Nile Red staining (literature guidance):

• Polarity and solvatochromism: Nile Red’s fluorescence is strongly enhanced in hydrophobic media and quenched in water. Working stocks are commonly prepared in polar aprotic or alcoholic solvents (e.g., DMSO, acetone, ethanol, isopropanol) and then diluted into aqueous buffers immediately before use so the dye partitions into lipid phases.
• Miscibility: DMSO/alcoholic stocks are miscible with aqueous buffers at low final percentages (typically ≤1% v/v) without precipitation, facilitating delivery to cells or tissues.
• Matrix effects: High water content around the dye reduces quantum yield and red-shifts emission. Keep organic cosolvent at the minimum that ensures delivery but avoids biological perturbation (e.g., ≤0.5–1% v/v, application-dependent).

When to choose alternatives:

• If solvent autofluorescence or toxicity is a concern, compare ethanol vs. isopropanol vs. DMSO carriers; background and cell compatibility can differ.
• For fixed samples, isopropanol/ethanol can aid lipid preservation; for live-cell imaging, DMSO at very low % is often preferred due to lower volatility.

Note: Because the specific solvent for this item is not disclosed here, validate compatibility with your sample type and instrument filters, and consult the SDS for solvent identity and handling.

Storage and Reconstitution

Item-specific conditions:

• Storage: Store at -20°C (provided). Protect from light.
• Shipment: Shipped in ice chest with ice pads (provided) to maintain low temperature.
• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Concentration: 100 μg/mL (provided). Ready-to-use solution; no reconstitution required.

Handling guidance:

• Keep tightly closed in an amber or foil-wrapped container to minimize light exposure.
• Avoid repeated freeze–thaw cycles; if frequent use is anticipated, aliquot upon first thaw into light-protected microtubes and refreeze at -20°C.
• Allow aliquots to equilibrate to room temperature briefly before opening to reduce condensation; mix gently before use to ensure homogeneity.
• Do not store working dilutions for extended periods unless stability has been verified for your solvent/buffer system; dye performance can degrade with light, oxygen, and moisture.

Stability/disposal:

• Shelf-life and stabilizers are Not specified for this item; refer to CoA/Spec Sheet.
• Dispose of unused solution and rinsates as hazardous chemical waste according to institutional and local regulations.

Research Use Only: For research use only (provided).

Structure and Identity

This product is a ready-to-use Nile Red Staining Solution at 100 μg/mL for fluorescence microscopy/flow cytometry of neutral lipids and lipid droplets.

• SKU: N1528845
• Product format: solution, 100 μg/mL (item-specific)
• Chemical identity of chromophore: Nile Red (general literature name; also called Nile Blue A oxazone, 9-diethylamino-5H-benzo[a]phenoxazin-5-one)
• Molecular formula: Not specified for this item; refer to CoA/Spec Sheet. (Literature for Nile Red provides a defined formula and MW, but item-level identifiers are not supplied here.)
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
• SMILES: Not specified for this item; refer to CoA/Spec Sheet.
• InChIKey: Not specified for this item; refer to CoA/Spec Sheet.
• CAS: Not specified for this item; refer to CoA/Spec Sheet.

Structural features (general, literature):

• Polycyclic, planar benzo[a]phenoxazone core with a dialkylamino substituent (electron-donating) conferring strong intramolecular charge transfer (ICT) character.
• Highly lipophilic; fluorescence is strongly environment- and polarity-dependent (solvatochromic probe). Fluorescence is quenched in protic/polar aqueous media and enhanced in hydrophobic phases.
• No stereogenic centers; the dye is achiral.

2D description (general): fused tricyclic/heteroaromatic framework containing an oxazine-type ring and carbonyl (lactam/quinone-like) functionality; para-dialkylamino phenyl moiety conjugated to the chromophore, enabling red-shifted absorption/emission in nonpolar media.

Synthetic Utility

Not typically applicable. Nile Red staining solutions are used as analytical/visualization reagents rather than as building blocks or reactants in organic synthesis.

Related analytical utility (literature):

• As a solvatochromic indicator, Nile Red can report on microenvironment polarity in reaction media, micelles, or polymer matrices, aiding optimization of formulations or reaction conditions involving surfactants/organogels.
• Can serve as a fluorescent tracer to monitor phase behavior, emulsification, or extraction processes, without participating chemically in the transformation.

Target Specificity

Not applicable. This product is a small-molecule fluorescent dye solution and is not an antibody or affinity reagent. No antigen/epitope or species specificity applies.