NBD-X PE , CAS No.N1450287

CAS: N1450287 Cat. No.: N1450287 Formula: C₅₅H₁₀₁N₆O₁₂P Peso molecolare: 1069.40

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Panoramica

NBD-X PE is a derivative of NBD-PE, where the NBD part is replaced by NBD-X acid.

Specifications

Condizioni di conservazione di stoccaggio

Store at -20°C

Spedito in

Ice chest + Ice pads

Questo prodotto richiede spedizione a catena fredda. I servizi di terra e altri servizi economici non sono disponibili.

Nomi e identificatori

• Peso molecolare: 1069.40

Application Protocols

Item-specific tested applications and recommended conditions: Not specified for this item; refer to CoA/Spec Sheet and SDS.

Note: Because no application data are provided for SKU N1450287, Aladdin does not offer validated protocols for this specific product. Users should consult peer-reviewed literature on NBD-PE handling and optimization for their chosen assay (e.g., liposome incorporation, flip-flop measurements, lipid transfer assays) and perform in-house method development and controls.

Biological Roles

Item-specific: None provided.

Literature/general perspective (for research context; not item-specific claims):

• Phosphatidylethanolamine (PE) is a major zwitterionic phospholipid in biological membranes, contributing to curvature, fusion, and membrane protein function. NBD-X PE acts as a fluorescent analog that mimics many aspects of native PE while enabling optical readouts.
• Membrane dynamics: NBD fluorescence is sensitive to polarity and quenchers, allowing assessments of local membrane order, leaflet asymmetry, and protein-induced remodeling.
• Trafficking pathways: NBD-PE analogs are used to study biosynthetic and endocytic routing of lipids, inter-organelle transport, and the activity of lipid flippases/floppases and scramblases in vitro and in cells.
• Enzyme substrates: Depending on acyl composition and headgroup accessibility, NBD-PE can serve as a substrate for phospholipases (A1/A2/D), enabling kinetic and mechanistic studies by fluorescence or HPLC detection.
• Protein interactions: The probe reports binding/insertion events for lipid-binding domains (e.g., C2, PH, START) by changes in intensity or anisotropy, facilitating quantitative affinity measurements.
• Microenvironment reporting: In less polar, tightly packed domains, NBD emission is brighter and blue-shifted; in disordered or aqueous-exposed regions, fluorescence diminishes—useful for domain mapping and phase separation studies.

Note: While NBD-PE is broadly informative, its photophysical and steric properties can alter native behavior; always include appropriate controls with unlabeled lipids and titrate probe mol%.

Buffer Applications

Item-specific: None provided.

General laboratory guidance for using NBD-X PE in buffers (not item-specific specifications):

• Aqueous dispersion: NBD-PE is amphiphilic and typically requires incorporation into lipid vesicles, mixed micelles, or protein complexes to remain dispersed in aqueous buffers. Common buffers include PBS, HEPES, or Tris at physiological ionic strength.
• Additives: 0.1–1% (w/v) nonionic surfactants (e.g., mild detergents) can solubilize NBD-PE for enzymatic assays; serum albumin can act as a carrier but may sequester the probe and alter fluorescence.
• pH range: The PE headgroup is zwitterionic across pH ~6–8; extreme pH can promote hydrolysis (acid/base-catalyzed) or affect fluorescence.
• Quencher-based assays: For flip-flop experiments, sodium dithionite (freshly prepared, oxygen-depleted) selectively reduces/quenches NBD on the outer leaflet; rigorous control of buffer oxygenation and pH (~7.4) is important.
• Light protection: Use amberware/foil during buffer prep and incubation; minimize exposure during measurements to limit photobleaching.
• Filtration: Filter buffers (0.22 µm) to reduce scattering background in fluorescence plate readers and microscopy.

Note: NBD-labeled PE is not a buffering agent itself; its use in buffers is as a fluorescent probe within lipid assemblies. Optimize buffer composition for your biological model and detection modality.

Green Alternatives

Context (general guidance; not item-specific):

• The primary environmental consideration with NBD-labeled lipids is solvent usage rather than the probe itself. Traditional dissolution for film hydration uses chlorinated solvents (CHCl3, DCM), which carry environmental and health burdens.

Greener handling strategies:

• Substitute ethanol or isopropanol for stock solutions where feasible, acknowledging longer drying times and potential residuals.
• Use ethyl acetate or 2-methyltetrahydrofuran (2-MeTHF) instead of DCM/THF for certain film-casting steps when solubility permits.
• Employ miniaturized, closed-vessel evaporation (rotavap with cold traps) to reduce emissions and capture solvents for recycling.
• Optimize probe loading (mol%) to minimize material and solvent use while maintaining adequate signal.

Comparison (literature-based; typical behavior):

• Chloroform vs ethanol:
  • Solubility: CHCl3 excellent; EtOH moderate.
  • Drying: CHCl3 fast; EtOH slower, but safer/greener.
  • Biological carryover: CHCl3 none if fully evaporated; EtOH tolerable at ≤0.1% v/v in many assays.
• THF vs 2-MeTHF:
  • Both dissolve amphiphiles; 2-MeTHF is bio-based, forms fewer peroxides, and can be a greener alternative.

Caveats:

• Verify probe solubility and absence of precipitation when switching to greener solvents.
• Ensure complete solvent removal before hydrating lipid films to avoid bilayer defects.

Always align with institutional green chemistry policies and waste segregation rules.

Pharmaceutical Uses

Item-specific: None provided.

General, non-clinical context (not item-specific and not therapeutic claims):

• NBD-X PE is a research-only fluorescent lipid probe and is not an approved excipient. However, in pharmaceutical R&D, such probes are used experimentally to study liposomal formulations, lipid nanoparticle (LNP) assembly, and drug–membrane interactions.
• Formulation development: Incorporation of trace amounts of NBD-PE into model membranes can aid in tracking bilayer integrity, cargo leakage, and fusion events during process optimization.
• Process analytics: Fluorescent lipids assist in monitoring mixing, extrusion, and size-homogeneity by microscopy or fluorescence spectroscopy.
• Regulatory status: No pharmacopeial monograph is associated with NBD-PE probes; materials are generally supplied for research use only. Any use in manufacturing or clinical settings would require separate qualification and is outside the scope of standard research reagents.

Recommendation: Restrict use to laboratory research and method development; consult quality/regulatory teams before any non-research application.

Physical Properties

Item-specific (from Product Data):

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight / formula: Not specified for this item; refer to CoA/Spec Sheet.

Literature/general values and behavior (for context; not item-specific specifications):

• Physical state: NBD-labeled phospholipids are typically yellow-green solids or foams, light-sensitive.
• Solubility: readily soluble in polar aprotic organics such as chloroform, dichloromethane, and THF; soluble in ethanol and isopropanol; sparingly soluble in water alone but dispersible in aqueous buffers when incorporated into liposomes or with detergents (e.g., 0.1–1% w/v nonionic surfactant).
• Partitioning: highly amphiphilic; strong partitioning into lipid bilayers and micelles; negligible partition into purely aqueous phases.
• Photophysics of NBD (literature, microenvironment-dependent):
  • Excitation λmax ~460–470 nm (commonly ~465 nm)
  • Emission λmax ~530–545 nm (commonly ~535 nm)
  • Quantum yield and emission intensity increase in less polar, non-aqueous environments; fluorescence is quenched by water and certain nucleophiles/thiols.
• Stability: NBD fluorophore is susceptible to photobleaching and nucleophilic attack; lipid esters may hydrolyze under strong base or acid; oxidation of unsaturated acyl chains can occur.
• Aggregation: forms bilayers or vesicles with other lipids; critical micelle/bilayer behavior depends on the matrix lipid composition and temperature (gel/fluid transition).

Important: The above properties are typical of NBD–PE probes and are provided for planning purposes only. Do not treat as Aladdin item specifications.

Quality and Grades

Item-specific (from Product Data):

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

Professional guidance on grades for fluorescent lipids (general; not item-specific):

• Research/biochemical grade: typically implies high purity and low autofluorescent contaminants; suitable for fluorescence microscopy, flow cytometry, and kinetic assays.
• HPLC-grade certification (when offered): indicates purification by preparative HPLC and narrow impurity profile—important for quantitative transport/flip-flop assays and for minimizing background fluorescence.
• Fatty acyl homogeneity: some vendors supply defined acyl chains (e.g., 16:0/6:0) versus mixtures; defined compositions aid reproducibility of membrane phase behavior.
• Counterion/salt form: presence of counterions (e.g., ammonium, sodium) and residual solvents can influence miscibility and bilayer insertion efficiency; check CoA for residual solvent and counterion content.
• Photometric specs: some listings provide absorbance/fluorescence criteria for NBD; absence of these requires in-house verification by excitation/emission scans after reconstitution.

Recommendations:

• Prior to critical experiments, verify identity and purity by analytical HPLC with fluorescence detection (Ex ~465 nm/Em ~535 nm) and/or MS.
• Record batch/lot numbers and retain CoA for method validation and reproducibility.

Reaction and Applications

Item-specific manufacturer applications: Not specified for this item.

Literature/general research applications of NBD-X PE (context; not item-specific claims):

• Membrane trafficking and endocytosis: incorporation into liposomes or cell membranes to visualize lipid internalization and vesicular transport by fluorescence microscopy or flow cytometry.
• Lipid transfer/transport assays: quantification of non-vesicular transfer between membranes mediated by lipid transfer proteins; monitor NBD fluorescence changes upon environment shift or FRET with complementary dyes.
• Flip-flop kinetics: assess transbilayer movement using fluorescence quenchers (e.g., dithionite reduces/quenches NBD in the outer leaflet) to resolve leaflet distribution over time.
• Enzymatic assays: substrates for phospholipase A2/A1 or phospholipase D—monitor release or transformation by HPLC-fluorescence.
• Phase behavior/probe partitioning: evaluate membrane order and domain formation; NBD’s polarity-sensitive fluorescence reports on local microenvironment.
• Calibration standards: define instrument settings for Ex ~465 nm/Em ~535 nm channels in imaging/plate readers.

Practical tips (general):

• Protect from light throughout handling; prepare fresh working dilutions.
• For cell work, preincorporate into carrier liposomes or use BSA-complexation to modulate delivery and reduce nonspecific adsorption.
• Confirm incorporation by thin-layer chromatography (TLC) with fluorescence detection or by extracting membranes and analyzing via HPLC-FLD.

Note: Optimize probe mol% (often 0.1–2 mol% of total lipid) to balance signal and minimal membrane perturbation.

Reaction Conditions

Item-specific: None provided.

General guidance for experimental use (not chemical synthesis; literature-based):

• Liposome incorporation:
  • Dissolve NBD-PE with bulk phospholipids in CHCl3 or EtOH; cast thin film; dry thoroughly; hydrate with appropriate buffer above the highest Tm of the lipid mixture; vortex/sonicate as needed; protect from light.
  • Typical probe loading: 0.1–2 mol% relative to total lipid for microscopy/transport assays; optimize empirically.
• Fluorescence measurements:
  • Excitation around 460–470 nm; emission collection around 530–545 nm. Use narrow-band filters to reduce background; verify instrument linearity with serial dilutions.
  • Temperature control improves reproducibility due to membrane phase transitions.
• Quenching/kinetics:
  • For flip-flop assays, add sodium dithionite to outer leaflet; maintain anaerobic conditions for stable quencher behavior. Record rapid time courses (sub-second to minutes) depending on system.
• Enzymatic assays:
  • Use mixed micelles (e.g., detergent/lipid) to present substrate; monitor fluorescence changes or analyze aliquots by HPLC-FLD.

Cautions:

• Avoid strong bases/acids and prolonged heating to prevent ester/amide hydrolysis.
• Minimize photobleaching by reducing excitation intensity and exposure time.

Note: These are general literature conditions for NBD–PE experiments and are not validated for this specific item.

Safety and Handling

Item-specific (from Product Data):

• GHS classification, pictograms, signal word, and H-statements: Not specified for this item; refer to the product SDS.
• Storage conditions: Store at −20°C.
• Shipping: Ice chest + ice pads.

General laboratory safety guidance (literature/practice; not a substitute for the SDS):

• PPE: wear lab coat, safety glasses, and appropriate chemically resistant gloves. Handle powders/films in a fume hood to minimize inhalation of dust or solvent vapors when preparing stocks.
• Light sensitivity: NBD fluorophores photobleach; protect material and solutions from light (amber vials, foil wrap).
• Incompatibilities: avoid strong bases/acids (ester/amide hydrolysis), strong nucleophiles (can react with NBD), and oxidants (lipid peroxidation). Avoid prolonged exposure to moisture and elevated temperatures.
• Solvent hazards: common dissolution solvents (e.g., CHCl3, DCM, THF, ethanol) have their own hazards—consult respective SDS; minimize exposure and use in ventilated enclosures.
• First aid (overview): in case of skin/eye contact, rinse with water for ≥15 min; if inhaled, move to fresh air; if ingested, rinse mouth—seek medical attention in all cases.
• Waste: collect organic solutions and contaminated materials as halogenated/non-halogenated organic waste per local regulations; avoid drain disposal.

Disclaimer: For authoritative hazard and response information, always consult the SDS for this specific catalog item.

Solvent Selection

Item-specific (from Product Data):

• No item-specific solvent recommendations provided; refer to CoA/Spec Sheet.

Literature/practical guidance for NBD-labeled phosphatidylethanolamines:

• Primary dissolution: good solubility in chloroform, DCM, and THF; moderate solubility in ethanol or isopropanol; DMSO can be used for aqueous dosing but may affect membrane properties above 0.5–1% v/v.
• Working stocks:
  • Organic stocks (1–10 mg/mL) in CHCl3 or EtOH for thin-film hydration and liposome preparation.
  • Aqueous delivery via ethanol or DMSO cosolvent: add small aliquots to vigorously stirred buffer containing lipid/detergent micelles to avoid precipitation.
• Miscibility/profile: amphiphilic; partitions strongly into lipid phases. In purely aqueous buffers, use with liposomes, detergents (e.g., Triton X-100 analogs), or cyclodextrins to ensure dispersion.
• When to choose alternatives:
  • If chlorinated solvents are restricted, use ethanol or isopropanol for stock solutions, accepting slower film formation and potential residual solvent in bilayers.
  • For highly sensitive cells, minimize DMSO/EtOH to ≤0.1–0.2% v/v in final media.

Quick comparison (general):

• CHCl3: excellent solubility; ideal for lipid film prep; hazardous/ozone-depleting concerns.
• DCM: rapid evaporation; good solubility; more volatile; halogenated waste.
• THF: good solvent; peroxide former—manage peroxides.
• EtOH/IPA: greener; acceptable solubility; slower drying; biological compatibility is better.

Note: Validate solvent choice empirically for your membrane system.

Storage and Reconstitution

Item-specific (from Product Data):

• Storage conditions: Store at −20°C.
• Shipping: Ice chest + ice pads.

General best practices for fluorescent lipid probes (literature/practical guidance; not item-specific specifications):

• Protect from light during storage and handling (amber vials, foil wrap).
• If supplied as a solid/film: equilibrate to room temperature in the dark before opening to minimize moisture condensation; promptly reseal under inert gas if possible.
• Reconstitution: prepare concentrated stocks in dry, oxygen-free organic solvent (e.g., CHCl3 or ethanol). For aqueous systems, introduce via cosolvent or after incorporation into lipid films; ensure complete removal of volatile solvents prior to hydration.
• Aliquoting: prepare single-use aliquots to avoid repeated freeze–thaw and light exposure.
• Stability: avoid prolonged exposure to high pH, strong nucleophiles, and oxidants. Store solutions at ≤−20°C when feasible; use freshly thawed aliquots promptly.
• Documentation: record lot number, preparation date, solvent, and concentration; verify by UV–Vis/fluorescence scan after preparation.

Reminder: In the absence of item-specific stabilizer or solvent data, consult the CoA/Spec Sheet and SDS for definitive instructions for SKU N1450287.

Structure and Identity

Item-specific (from Product Data):

• Product name: NBD-X PE (SKU: N1450287)
• CAS: N1450287 (catalog placeholder)
• 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 / InChIKey: Not specified for this item; refer to CoA/Spec Sheet.

Literature/general description (for context; not item-specific):

• NBD-X PE commonly denotes a phosphatidylethanolamine (PE) lipid bearing an NBD (7-nitrobenz-2-oxa-1,3-diazol-4-yl) fluorophore attached to the PE headgroup via an aminohexanoyl ("X"; 6-carbon) spacer. The hydrophobic tails are typically long-chain fatty acyls (e.g., C16–C18), which may vary by manufacturer.
• Functional groups/features: glycerol backbone esterified to two fatty acids (sn-1, sn-2), phosphate linking to ethanolamine; the ethanolamine nitrogen is acylated with the NBD–(CH2)6–CO linker. Contains an anionic phosphate and cationic ammonium at physiological pH (zwitterionic), plus the strongly electron-withdrawing NBD nitro-aza-benzofurazan ring.
• 2D structure (verbal): a diacylglycerol tail domain (two long alkyl chains) connects through a phosphate to an ethanolamine headgroup; the headgroup nitrogen bears an amide to a hexanoyl spacer terminating in an NBD aromatic heterocycle with a nitro substituent and embedded oxygen/nitrogen atoms.

Notes:

• Exact fatty acyl composition, stereochemistry (sn-1/sn-2), and salt form can differ by source. Consult the product’s CoA for definitive identity data.

Synthetic Utility

Item-specific: None provided.

General perspective (not item-specific):

• NBD-X PE is a terminally functionalized lipid probe intended for biophysical/biological assays rather than as a synthetic building block. The NBD chromophore is already installed on the PE headgroup via an amide linkage to a hexanoyl spacer.
• Limited reactivity: The molecule contains hydrolyzable esters (acyl chains) and an amide (linker to NBD). Under standard synthetic conditions, these groups are labile, making the compound unsuitable as a substrate for further derivatization without damaging the probe.
• Utility in assembly: Its primary “synthetic” role is in supramolecular assembly—co-formulation into liposomes, supported lipid bilayers, or nanodiscs—where it serves as a functional component to report on structure and dynamics.
• Analytical utility: Can be used as a fluorescent standard in TLC/HPLC method development for lipid separations, given its distinct excitation/emission.

If chemical modification is desired, it is usually performed on precursor lipids (e.g., PE with protected amine) prior to NBD installation or by using alternative headgroup chemistries (azide/alkyne handles) for click ligations.

Target Specificity

Item-specific: Not applicable—this product is a small-molecule fluorescent lipid probe, not an antibody or targeted biologic.

No target/epitope, species reactivity, clone, or isotype information is provided or applicable for this item.