N-Hexanoyl-NBD-lactosylceramide - ≥98%

Cat. No.: H275573

GRADE & PURITY ≥98%

Storage

Store at -20°C,Desiccated

Shipped In

Ice chest + Ice pads

Descripción general

Store at -20°C. Store under desiccating conditions. The product can be stored for up to 12 months.

Specifications

Especificaciones y pureza

≥98%

Condiciones de almacenamiento de almacenamiento

Store at -20°C,Desiccated

Enviado en

Ice chest + Ice pads

Este producto requiere envío en cadena de frío. Los servicios terrestres y otros servicios económicos no están disponibles.

Pureza

≥98%

Application Protocols

Item‑specific validated protocols are not provided in the Product Data. The following literature‑style templates are commonly used with NBD‑labeled glycosphingolipids; optimize for your system.

1. Stock solution preparation

• Dissolve at 1–5 mM in dry DMSO or CHCl3:MeOH (1:1) under low light. Aliquot into amber vials; purge with N2/Ar; store at −20 °C, desiccated.

2. Cell membrane labeling (live cells)

• Prepare a 1–10 µM working solution in pre‑warmed HBSS or culture medium containing 0.5–1% fatty‑acid‑free BSA (final DMSO ≤0.1% v/v if using DMSO stock).
• Incubate cells 5–20 min at 37 °C in low light. Wash 3× with buffer. Image promptly (Ex ~470–480 nm, Em ~530–540 nm).

3. Liposome incorporation

• Co‑dissolve matrix lipids and probe in CHCl3/MeOH, form thin film, dry thoroughly, hydrate with buffer above Tm, vortex and extrude. Typical doping: 0.1–1 mol%.

4. TLC analysis of enzymatic reactions

• Quench reaction with CHCl3:MeOH, spot on silica TLC. Develop in CHCl3:MeOH:Water 65:25:4 (v/v/v). Visualize under blue light; quantify by densitometry.

Controls and cautions

• Include unlabeled LacCer controls to assess label effects. Protect from light at all times. Validate cell viability for solvent/carrier concentrations used. Sterile‑filter aqueous working solutions when applicable.

Biological Roles

Literature/general context:

• Lactosylceramide (LacCer) is a central glycosphingolipid intermediate in the biosynthesis of more complex gangliosides and globosides. It resides primarily in the outer leaflet of the plasma membrane, contributing to membrane organization, microdomain (raft) formation, and cell–cell interactions.
• Enzymatic pathways: LacCer is generated from glucosylceramide and is a substrate for glycosyltransferases (e.g., addition of galactose/N‑acetylgalactosamine to form globosides) and sialyltransferases (toward gangliosides). It is also catabolized by glycosidases in lysosomes.
• Signaling interfaces: Through its carbohydrate headgroup, LacCer can engage lectins and modulate adhesion and receptor clustering. Its ceramide backbone influences bilayer order and curvature.
• NBD fluorophore: Acts as an environment‑sensitive reporter. Upon insertion into membranes, NBD fluorescence typically increases and blue‑shifts relative to aqueous environments. This enables monitoring of trafficking, phase partitioning, and enzymatic conversion.
• Cellular uptake/trafficking: Short‑chain (C6) NBD‑glycosphingolipids are commonly internalized and routed to Golgi or endolysosomal compartments, allowing visualization of metabolism and sorting in live cells (pathway dependence varies by cell type and conditions; literature observations).

Note: The above describes general biological roles of LacCer and NBD‑labeled analogs. This product is provided strictly for research use; no clinical or diagnostic use is implied.

Buffer Applications

This compound is not a buffering agent and does not establish defined pH ranges.

Practical buffer guidance for using NBD‑lactosylceramide in biological assays (literature/general):

• Hydration/dispersion: Use HEPES‑buffered saline (HBS) or PBS supplemented with 0.1–1% fatty‑acid‑free BSA to disperse the lipid and prevent aggregation.
• Detergent‑assisted delivery: Low concentrations of nonionic detergents (e.g., 0.01–0.1% Triton X‑100 or Tween 20) can aid solubilization; ensure compatibility with your cells/assay.
• pH: Maintain near‑neutral pH (7.2–7.4) to minimize hydrolysis of glycosidic and amide bonds and to preserve NBD fluorescence.
• Filtration: 0.22 µm filtration of final working solutions can remove particulates; avoid strong pressure that could shear micelles.

For buffer recipes and volumes, consult standard cell culture or membrane biophysics protocols. For chromatographic or enzymatic assays, refer instead to the Reaction & Applications and Application Protocols sections.

Green Alternatives

Because workflows with this probe often rely on halogenated solvents, greener process choices can reduce environmental and health burdens without compromising data quality.

Greener solvent considerations (literature/general):

• Replace CHCl3 with EtOAc, IPA, or MeOH where feasible (e.g., for film dissolution prior to final chloroform rinse, or for cleaning glassware). Note: EtOAc/MeOH may not solubilize concentrated lipid films as efficiently.
• Consider CPME or 2‑MeTHF for some dissolution/transfer steps; these ethers have better green metrics than CH2Cl2, but check solubility—glycosphingolipids may require alcohol cosolvents.
• Use DMSO for stock solutions when biological compatibility is paramount, limiting final DMSO to ≤0.1% v/v in cell media (literature guidance).
• Minimize solvent volumes through concentrated stocks, positive‑displacement pipetting, and nitrogen‑assisted thin‑film formation.

Comparison (typical attributes):

• CHCl3: excellent solubilizer; poor EHS profile; halogenated waste stream.
• CH2Cl2: volatile, effective; also halogenated waste; higher vapor exposure risk.
• EtOAc/IPA/MeOH: lower toxicity, non‑halogenated waste; may need higher volumes or heat.
• DMSO: high solvency, benign; hygroscopic and can carry compounds through skin—use gloves.

Balance and trade‑offs:

• For quantitative lipid blending, small amounts of CHCl3/MeOH may still be required; offset impact by microscale handling, cold traps, and rigorous waste segregation.
• Adopt amber glassware and low‑light workflows to extend probe lifetime, reducing resynthesis/reorders and overall environmental footprint.

Pharmaceutical Uses

No pharmacopeial or excipient designation is provided for this item. It is supplied for research use only.

Relevant research/manufacturing‑adjacent uses (literature/general):

• Analytical reference standard: Employed as a fluorescent internal or external standard in development/qualification of lipid extraction, TLC/HPLC separations, and detector settings in QC‑like research environments.
• Process visualization: Useful for tracing lipid handling steps in formulation research (e.g., tracking incorporation into liposomes or nanoparticles) without implying any therapeutic role.
• Method development: Optimization of extraction recoveries from complex matrices, detector linearity, and limit of detection/quantitation for glycosphingolipid assays.

No claims are made regarding therapeutic efficacy, diagnostic performance, or suitability for human or veterinary use. For any regulated application, consult relevant compendia and perform full in‑house qualification.

Physical Properties

Item‑specific physicochemical specifications are not provided in the Product Data and should be confirmed on the CoA/Spec Sheet.

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
• Melting point, boiling point, density, refractive index: Not specified for this item; refer to CoA/Spec Sheet.

Literature/general characteristics for NBD‑labeled C6‑lactosylceramide:

• Physical state: typically a light‑sensitive, amorphous to waxy solid or film at room temperature when solvent is evaporated.
• Amphiphilic behavior: forms micelles or inserts into lipid bilayers; tends to aggregate in aqueous buffers without carrier proteins or detergents.
• Solubility profile (qualitative):
  • Good solubility in CHCl3/MeOH mixtures (e.g., 2:1 to 1:1, literature), CH2Cl2, and DMSO.
  • Limited aqueous solubility; can be dispersed using BSA, cyclodextrins, or nonionic detergents (e.g., 0.05–0.1% Triton X‑100 or Tween 20, literature guidance).
• Spectroscopic properties (NBD, literature):
  • Absorption maxima typically ~465–480 nm; emission ~530–545 nm, with polarity‑dependent shifts and quantum yield changes.
  • Pronounced photobleaching under intense illumination; protect from light.

Practical notes:

• Prepare stock solutions under low light in dry, oxygen‑free organic solvents when possible.
• Avoid prolonged exposure to aqueous buffers without carriers to minimize aggregation and fluorescence quenching.

Quality and Grades

• Grade/Purity: Not specified for this item; refer to CoA/Spec Sheet.

Context for this product class (literature/general):

• Purity expectations: Fluorescent glycolipids intended for imaging/biochemistry are commonly supplied at high purity (often ≥95% by HPLC). Verify the certificate for exact purity, identity confirmation (HRMS, 1H/13C NMR), and residual solvent levels.
• Identity controls: For lipids, suppliers often confirm lactose headgroup integrity (enzymatic or NMR), ceramide chain composition, and presence of NBD (UV‑Vis/fluorimetry). Co‑chromatography with standards and MS fragmentation can corroborate structure.
• Stabilizers: Typically none are added; protection from moisture and light is used instead. If any stabilizer or antioxidant is present, it will be declared on the CoA—none is specified here.
• Fluorescence QC: Batch‑to‑batch comparison of excitation/emission spectra, quantum yield in standard solvent (e.g., MeOH) and stability under storage may be provided by some manufacturers; consult the Spec Sheet.
• Endotoxin/bioburden: Not applicable to small‑molecule research reagents unless explicitly stated.

What is specific to this item:

• Storage guidance is defined: Store at −20 °C, desiccated; shipped cold. This is consistent with maintaining fluorophore integrity and minimizing hydrolysis of glycosidic and amide bonds.
• For critical applications (quantitative trafficking assays), we recommend documenting lot number, retesting purity by analytical HPLC, and recording fluorescence spectra upon receipt.

Reaction and Applications

This product is primarily a functional probe, not a synthetic reagent. Key research applications (literature/general) include:

• Membrane trafficking and endocytosis studies: NBD‑lactosylceramide integrates into the plasma membrane and undergoes internalization via clathrin‑independent pathways; fluorescence enables tracking of intracellular routing to Golgi/lysosomes depending on cell type and conditions.
• Glycosphingolipid metabolism assays: As a substrate/traceable reporter for glycosyltransferases, glycosidases, and ceramidases; product formation can be analyzed by TLC or HPLC with fluorescence detection.
• Lipid raft/microdomain interrogation: Partitioning into ordered domains of model membranes; used to study phase behavior and protein–lipid interactions.
• Lipidomics standards: Spike‑in control for method development and recovery assessments in extraction and separation workflows.
• Model membrane reconstitution: Doping into liposomes or supported bilayers for single‑vesicle and microscopy studies.

Practical tips:

• Work under low light; NBD photobleaches and is polarity‑sensitive—fluorescence increases in nonpolar environments.
• Avoid prolonged exposure to basic conditions; β‑elimination or amide hydrolysis can reduce signal.
• For TLC visualization, excite near 470–490 nm and detect green emission; run standards alongside to monitor enzymatic conversions (e.g., to globotriaosylceramide or monosaccharide cleavage products).
• For cellular dosing, pre‑complex with BSA or deliver via cyclodextrins to minimize nonspecific precipitation and ensure reproducible uptake (literature practice).

Reaction Conditions

No item‑specific reaction conditions are defined because this product is intended as a fluorescent probe rather than a reagent. The following general, literature‑based conditions pertain to common experimental uses:

• Thin‑film hydration for liposomes: Dissolve matrix lipids and NBD‑lactosylceramide in CHCl3/MeOH (2:1), form a thin film under N2, desiccate ≥2 h, hydrate with HEPES‑buffered saline above the highest Tm of matrix lipids, then extrude. Typical dopant levels: 0.1–1 mol% probe (literature practice).
• BSA complex formation: Evaporate an aliquot to a thin film, add warm buffer containing 0.5–1% fatty‑acid‑free BSA, vortex/sonicate briefly in low light; use promptly.
• TLC separation (analytical): Silica gel; mobile phases such as CHCl3:MeOH:Water 65:25:4 (v/v/v) or 60:35:8 (literature examples). Visualize under 470–490 nm excitation; detect green emission.
• HPLC (analytical): Normal‑phase or HILIC with fluorescence detection (Ex ~470–480 nm, Em ~530–540 nm, literature). Gradient and column choice depend on matrix lipids.
• Cell labeling: Working concentrations often in the 0.5–10 µM range with carrier protein; incubate 5–30 min at 37 °C, then wash (literature practice). Optimize to minimize background and phototoxicity.

These conditions are guidance only; optimize for your system. Avoid strong base/acid, prolonged light exposure, and elevated temperatures that may hydrolyze the glycosidic or amide linkages or bleach NBD.

Safety and Handling

Hazard classification details are not provided in the Product Data and should be obtained from the item‑specific SDS.

• Signal word, H‑statements, GHS classes, pictograms: Not specified for this item; refer to SDS.

General laboratory safety guidance for NBD‑labeled glycolipids (literature/general):

• PPE: laboratory coat, safety glasses, and appropriate chemically resistant gloves (e.g., nitrile). Handle in a fume hood when using volatile organic solvents (CHCl3, CH2Cl2, MeOH).
• Light sensitivity: NBD fluorophore is photolabile. Minimize light exposure during handling and storage (use amber vials/foil wrapping).
• Incompatibilities: strong oxidizers, strong acids/bases (can hydrolyze glycosidic/amidic bonds), and reducing agents that may affect the nitroaromatic moiety.
• Peroxide concerns: not a typical peroxide former; however, many organic solvents used with this product can form peroxides—monitor solvent quality as appropriate.
• First aid (overview; follow SDS):
  • Inhalation: move to fresh air; seek medical attention if symptoms persist.
  • Skin/eye contact: rinse with water for ≥15 min; remove contaminated clothing; seek medical attention as needed.
  • Ingestion: rinse mouth; do not induce vomiting; seek medical attention.
• Spill response: absorb organic solutions with inert material; collect for disposal per institutional and local regulations.
• Waste: dispose of solutions and contaminated materials as hazardous chemical waste; segregate halogenated from non‑halogenated solvent waste.

Always defer to the SDS and institutional EHS policies for authoritative guidance.

Solvent Selection

This amphiphilic fluorescent glycolipid dissolves well in moderately polar organic solvents and mixed systems; it aggregates in pure water. Selection depends on the downstream application.

Polarity/miscibility (literature/general):

• Preferred stock solvents: CHCl3/MeOH (2:1 to 1:1), CH2Cl2, or dry DMSO for convenience and stability.
• Dispersants for aqueous work: BSA complexation, cyclodextrins, or nonionic detergents (e.g., 0.05–0.1% Triton X‑100) to prevent aggregation and enhance delivery to cells or vesicles.
• Liposome incorporation: dissolve lipid mixture in CHCl3/MeOH, form a thin film, dry thoroughly, then hydrate with buffer above Tm of matrix lipids; the probe partitions into bilayers.

When to choose each:

• DMSO stocks (1–5 mM): convenient for small‑volume dosing into cell media (<0.1% v/v final DMSO to protect cells; literature practice). Protect from light.
• CHCl3/MeOH: ideal for precise lipid blending and TLC/HPLC analytics; evaporates cleanly to give films for liposome prep.
• Aqueous buffer with carrier: for immediate biological assays; prepare fresh to minimize hydrolysis/photodegradation.

Comparison (considerations):

• CHCl3 vs CH2Cl2: CHCl3 solubilizes glycosphingolipids well but is less green; CH2Cl2 is more volatile; both require fume hood.
• IPA/MeOH cosolvents: greener but may require larger volumes or carriers to prevent precipitation.

Always confirm compatibility of solvent residues with your cells/assays and filter sterilize aqueous dispersions when needed.

Storage and Reconstitution

• Storage conditions (item‑specific): Store at −20 °C, desiccated. Protect from light. Avoid repeated freeze–thaw.
• Shipping (item‑specific): Shipped in an ice chest with ice pads to maintain cold chain.

Best practices (literature/general):

• Packaging: Keep in amber glass vials with PTFE‑lined caps. After opening, immediately reseal under dry inert gas (N2/Ar) if available to limit moisture and oxygen exposure.
• Stability: NBD fluorophores can photobleach and may slowly degrade under moisture/heat. Maintain low temperature and desiccation; avoid prolonged storage at room temperature.
• Reconstitution:
  • For organic stocks: Dissolve in dry DMSO or CHCl3:MeOH (1:1 to 2:1). Vortex gently; verify clarity. Prepare small aliquots (e.g., 10–50 µL) to minimize freeze–thaw.
  • For aqueous use: Formulate immediately before use by diluting an organic stock into buffer containing carrier (e.g., 0.5–1% fatty‑acid‑free BSA) with vigorous mixing; alternatively, incorporate into liposomes via thin‑film hydration.
• Working solution handling: Keep on ice and protected from light; use within the same day. Do not store diluted aqueous solutions for extended periods.

For any lot‑specific stability or solubility data, consult the CoA/Spec Sheet and the SDS.

Structure and Identity

N‑Hexanoyl‑NBD‑lactosylceramide is a fluorescent glycosphingolipid probe: a lactosylceramide (Galβ1→4Glcβ1→1′Cer) bearing a short C6 acyl chain functionalized with an NBD (7‑nitrobenz‑2‑oxa‑1,3‑diazol‑4‑yl) fluorophore via an aminohexanoyl spacer (literature description). The molecule combines a hydrophobic ceramide backbone with a hydrophilic disaccharide headgroup and a neutral, environment‑sensitive NBD tag.

• SKU: H275573
• CAS: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular formula: Not specified for this item; refer to CoA/Spec Sheet.
• 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.

Structural features (literature/general):

• Glycolipid architecture: ceramide (sphingoid base amide‑linked to a fatty acyl chain) with a terminal lactose (Gal‑Glc) headgroup.
• Fluorophore: NBD ring (nitrobenzoxadiazole), typically attached at the ω‑end of a 6‑carbon acyl spacer (C6) on the ceramide amide nitrogen.
• Amphiphilicity: packs into membranes and liposomes; lactose headgroup confers aqueous interfacial localization, ceramide tail embeds in bilayers.
• 2D depiction in words: a disaccharide head connects via β‑glycosidic linkage to C1 of the ceramide; the ceramide amide bears a hexanoyl chain whose terminal amino group is coupled to NBD.

Notes:

• Exact stereochemistry and sphingoid base composition may vary by supplier specification; consult the CoA for definitive structural identifiers of this lot.

Synthetic Utility

As supplied, N‑Hexanoyl‑NBD‑lactosylceramide is the end product and is not typically a building block for further chemical synthesis in routine workflows. Nevertheless, it has utility in synthetic/biocatalytic method development (literature/general):

• Enzymatic transformations: Serves as a fluorescent substrate to benchmark glycosyltransferases (extension to higher glycosphingolipids), sialyltransferases, and glycosidases; product profiles are monitored by TLC/HPLC with fluorescence detection.
• Lipid remodeling assays: Ceramidases and acyltransferases can process the ceramide moiety; the NBD label facilitates kinetic studies and comparative enzyme profiling.
• Click/derivatization at the headgroup: While the lactose is not typically modified on this probe, selective oxidation (e.g., NaIO4 at C2–C3 diol) followed by oxime/hydrazone ligation has been explored on related glycolipids to append affinity tags—perform with caution to avoid NBD degradation (literature precedent).
• Partitioning standards: In model membranes, acts as a tracer to calibrate phase preference and leaflet distribution for new synthetic lipids or polymers.

If chemical modification is intended, confirm stability of the NBD fluorophore (sensitive to strong nucleophiles, bases, and reducing agents) and monitor by LC‑MS and UV‑Vis during process development.

Target Specificity

This product is not an antibody or targeted biologic; no antigen/epitope specificity is applicable.

Literature/general notes on functional partitioning:

• NBD‑lactosylceramide exhibits physicochemical “specificity” for lipid bilayers, enriching in membrane regions with appropriate packing and hydrogen‑bonding capacity (e.g., glycosphingolipid‑rich microdomains). It can serve as a substrate for enzymes that naturally process lactosylceramide, enabling pathway‑selective readouts.
• Any apparent protein targeting in cells (e.g., lectin interactions via the lactose headgroup) is context‑dependent and not intrinsic specificity of this reagent. Users should validate localization with orthogonal markers and appropriate controls.