NBD-Fructose , CAS No.940961-04-6

CAS: 940961-04-6 Cat. No.: N1452088 Formula: C₁₂H₁₄N₄O₈ Peso molecolare: 342.26 PubChem CID: 125181376

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Panoramica

NBD-Fructose is a fluorescent derivative of Fructose that is formed by coupling NBD-chloride with the amine group of amino fructose (EX/Em=472/538 nm).

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

• Sorrisi canonici: C1=C(C2=NON=C2C(=C1)[N+](=O)[O-])NCC(=O)C(C(C(CO)O)O)O
• IUPAC Name: (3S,4R,5R)-3,4,5,6-tetrahydroxy-1-[(4-nitro-2,1,3-benzoxadiazol-7-yl)amino]hexan-2-one
• InChIKey: PYUWRKIMILVYHE-GGZOMVNGSA-N
• INCHI: 1S/C12H14N4O8/c17-4-8(19)12(21)11(20)7(18)3-13-5-1-2-6(16(22)23)10-9(5)14-24-15-10/h1-2,8,11-13,17,19-21H,3-4H2/t8-,11-,12-/m1/s1
• PubChem CID: 125181376
• Peso molecolare: 342.26

Application Protocols

Item-specific tested applications are not provided for this product. The following are general, literature-based procedures for NBD-labeled small molecules (adapt to your system; not item-specific specs):

A. Preparation of stock and working solutions

• Weigh material quickly under low light. Dissolve to 10 mM in anhydrous DMSO. Vortex until clear; protect from light. Aliquot into amber tubes and store at −20 °C.
• For assays, dilute to 0.1–5 µM in HEPES-buffered saline (HBS) or PBS; keep final DMSO ≤0.5%.

B. Plate-based binding or uptake assay (fluorescence readout)

• Add 100 µL of probe working solution to wells containing target protein/cells.
• Incubate 5–60 min at desired temperature.
• Read fluorescence (Ex 465 ± 10 nm; Em 535 ± 15 nm). Optimize gain and background subtraction using buffer-only and solvent controls.
• Specificity control: co-incubate with 10–100× unlabeled fructose; quantify signal reduction.

C. Fluorescence spectroscopy

• Record excitation and emission spectra in the exact assay buffer to determine λmax and Stokes shift. Validate linearity over your concentration range.

D. Microscopy (optional)

• Use FITC/GFP filter sets as a starting point; verify bleed-through. Minimize light exposure to reduce photobleaching.

All procedures are for research use only; confirm suitability in pilot experiments.

Biological Roles

Item-specific note

• This product is for research use only.

General biology (fructose; literature)

• Fructose is a ketohexose central to carbohydrate metabolism, feeding into glycolysis via fructokinase and aldolase pathways. It is transported by facilitative transporters (e.g., GLUT5) and can interact with lectins in certain organisms.

Relevance of NBD labeling (literature/general)

• NBD-fructose functions as a fluorescent surrogate to probe recognition events where fructose participates. The compact NBD dye often preserves some binding features while providing optical readout. Because site/linker details for this item are not specified, users should confirm that the conjugate retains affinity for the intended target (transporter, enzyme, or lectin).
• Environment-sensitive fluorescence: NBD intensity typically increases in nonpolar or protein-bound environments, enabling detection of binding or membrane translocation as changes in signal rather than absolute concentration.

Use cases (research)

• Kinetic assays of hexose uptake in cells or vesicles using fluorescence plate readers.
• Binding studies with purified lectins or enzymes using fluorescence anisotropy/polarization.
• Microscopy of probe localization under live or fixed-cell conditions after toxicity and background controls.

Important caveat

• NBD labeling can alter transport/enzyme kinetics versus native fructose; benchmark with competition by unlabeled fructose to confirm biological relevance.

Buffer Applications

Applicability: NBD-Fructose is not a buffering reagent. However, buffer choice critically impacts fluorescence and biological assays.

General guidance (literature)

• Common assay buffers: PBS (pH 7.2–7.4), HEPES (pH 7.2–7.5), or Tris (pH 7.4–8.0). HEPES is often preferred for fluorescence due to low autofluorescence in the green channel; PBS is widely compatible with cells.
• pH effects: NBD fluorophores exhibit environment- and sometimes pH-dependent fluorescence; avoid extremes (pH <6 or >8) unless validated. Tris can show background at some wavelengths; verify with blanks.
• Organic cosolvent: 0.1–1% DMSO or ethanol can aid solubility; confirm that final solvent percentage does not perturb cells or proteins.
• Additives: 0.01–0.05% nonionic surfactant (e.g., Tween-20) may reduce adsorption to plastics; confirm that surfactant does not quench fluorescence.

Practical steps

• Prepare matched blanks containing the same buffer and cosolvent for baseline subtraction.
• Record excitation/emission spectra of working solutions in the exact buffer used for assays to establish optimal filter sets and minimize inner-filter effects.

Green Alternatives

Context: Fluorescent carbohydrate probes inherently require synthetic organic chemistry and aromatic fluorophores. “Greener” choices center on solvent selection, probe dose minimization, and choosing dyes with better photostability to reduce waste.

Potential alternatives/complements (literature; not item-specific)

• Alternative fluorophores: Coumarin- or fluorescein-labeled fructose derivatives may offer higher aqueous brightness, potentially reducing required probe concentration. Tradeoff: larger dye may perturb binding more than compact NBD.
• Solvent strategies: Replace DMF with DMSO, ethanol, or water/ethanol mixtures when compatible. Use minimal organic co-solvent (≤1% v/v) in biological assays.
• Energy and waste: Prepare concentrated stocks to minimize container use; employ amber, reusable glassware.

Comparison snapshot (general)

• NBD-fructose: Small, environment-sensitive; exc ~465 nm / em ~535 nm (literature). Brightness lower in water; often needs modest organic cosolvent.
• Fluorescein-fructose: Higher aqueous brightness; exc ~490 nm / em ~515 nm; more pH-sensitive around neutral pH.
• Coumarin-fructose: UV-blue excitation; may require specialized optics; good photostability in some cases.

Greener practice tips

• Optimize optical setup (bandpass filters, detector gain) to work at lower probe concentrations.
• Validate adsorption losses to plastics; switch to low-binding consumables to avoid repeated preparations.

Pharmaceutical Uses

This product is not intended for pharmaceutical use. No pharmacopeial status, excipient role, or clinical application is provided. For research use only.

General note

• Fluorescently labeled sugars, including NBD–carbohydrate conjugates, are used in research and development settings to study transport, binding, and formulation behavior but are not standard pharmaceutical excipients. Any use in manufacturing or regulated settings would require dedicated qualification and stability studies beyond the scope of this listing.

Physical Properties

Item-specific specs

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Purity/Grade: 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 properties for NBD fluorophores and NBD-labeled sugars (non-spec, for guidance only)

• Optical properties (NBD core, solvent-dependent):
  • Absorption λmax typically ~460–470 nm; emission λmax typically ~530–550 nm, with fluorescence strongly environment/polarity sensitive (literature).
  • Molar absorptivity for NBD dyes often 20,000–25,000 M−1 cm−1 near λmax (literature; varies with conjugate and solvent).
• Solubility: NBD–sugar conjugates are generally soluble in polar organic solvents (DMSO, DMF, MeOH) and may be water-dispersible with co-solvent or mild base; exact solubility for this item is not specified and should be verified empirically.
• Partitioning: NBD increases hydrophobicity versus native fructose; overall amphiphilicity depends on linker and substitution site (literature).
• Stability: NBD fluorophores are light-sensitive; avoid prolonged exposure to light. Hydrolytic stability depends on linkage chemistry (oxime/amine/ether), which is not specified for this item.

Do not interpret the above as specifications. For definitive values, consult the item’s CoA/Spec Sheet and SDS.

Quality and Grades

Item-specific quality information

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

How to interpret grade/purity for fluorescent carbohydrate probes (general guidance)

• Research grade: Typically indicates suitability for biochemical assays and imaging; may not include pharmacopeial testing.
• HPLC grade/assay purity: For fluorescent probes, high assay purity reduces background fluorescence from aromatic impurities; vendors may report HPLC area % and optical purity if relevant. If UV cutoffs or residual solvents are critical for your application, consult the CoA.
• Stabilizers: Not indicated for this item. Some NBD reagents include stabilizers or are supplied in amber vials to limit photobleaching. Absence/presence of stabilizers affects baseline fluorescence and long-term stability.
• Batch documentation to request (best practice):
  • HPLC/UPLC chromatogram and purity method.
  • Residual solvent profile (GC), water content (KF) if applicable.
  • Identity confirmation: HRMS, 1H/13C NMR; fluorescence spectrum.
  • Extinction coefficient reference conditions, if reported.

Practical tips

• Run an incoming QC check matching your assay: record absorbance/fluorescence spectra in your assay buffer to define λmax and intensity; compare lots.
• If quantitative binding/transport studies are planned, standardize concentration by weight-to-volume in dry solvent (e.g., DMSO) and verify by UV–Vis using an extinction coefficient measured in the same solvent/buffer.

Reaction and Applications

Focus: NBD-Fructose is primarily a fluorescent probe for biochemical/biophysical studies rather than a synthetic reagent.

Representative research applications (literature; expand-able)

• Transport/uptake studies: Probe for facilitated diffusion via hexose transporters or fructose-specific transport systems in cells, vesicles, or proteoliposomes. Fluorescence enables kinetic assays (e.g., stopped-flow, plate-based readouts).
• Lectin/glycan-binding analysis: Monitor binding to fructose-recognizing lectins using fluorescence polarization, anisotropy, or microplate binding assays.
• Enzymology: Substrate or reporter for fructokinase/isomerase pathways where labeling does not abolish recognition; verify activity empirically.
• Imaging: Visualization of probe accumulation or membrane association (NBD is environment-sensitive; brightness increases in hydrophobic domains).

Assay design considerations (general guidance)

• Validate that labeling position/linker preserves biological recognition; the exact regiochemistry for this item is not provided, so pilot experiments are essential.
• Calibrate fluorescence vs concentration in your assay buffer to account for inner-filter effects and environment-dependent quantum yield.
• Control experiments: unlabeled fructose competition; solvent controls (DMSO%), photobleaching curves; if using cells, viability controls.

Analytical methods

• Spectrofluorometry (typical λex ~465 nm; λem ~530–550 nm, literature).
• HPLC or LC–MS to confirm integrity in biological matrices; monitor for hydrolysis of the linker if applicable.

Reaction Conditions

This product is not primarily used as a reagent in chemical transformations. The following guidance addresses typical conditions for assays employing NBD-labeled small molecules (general literature; not item-specific specs):

• Stock solutions: Prepare in anhydrous DMSO at 1–10 mM; vortex and, if needed, briefly sonicate. Filter through 0.2 µm PTFE for particulate removal. Protect from light.
• Working solutions: Dilute into buffer (e.g., HEPES or PBS) to low micromolar range (0.05–10 µM) depending on detector sensitivity. Keep final DMSO ≤0.5–1% v/v for cell-based assays.
• Fluorescence measurement: Typical NBD settings λex ≈ 460–470 nm; λem ≈ 530–550 nm. Optimize excitation/emission slit widths and gain to avoid saturation.
• Temperature: Conduct binding/transport assays at the biologically relevant temperature (e.g., 25 °C for in vitro protein assays; 37 °C for mammalian cell assays), monitoring for photobleaching.
• Controls: Include solvent blanks, unlabeled fructose competition, and photostability time courses.

Note: If the intended use involves chemical coupling or surface immobilization, consult the item’s CoA to determine whether a reactive functional group is present; conditions will depend on that chemistry (e.g., amide coupling, oxime formation, click reactions).

Safety and Handling

Item-specific hazards

• GHS classification, pictograms, signal word, and H-statements: Not specified for this item; refer to SDS.

General safety guidance (literature/standard lab practice)

• Potential hazards: NBD fluorophores are nitroaromatic heterocycles; handle as harmful if swallowed, inhaled, or in contact with skin/eyes. Dust or powder may cause respiratory/eye irritation. Carbohydrate portion is low hazard, but the NBD moiety governs risk profile.
• PPE: Use lab coat, safety glasses, and appropriate chemically resistant gloves (e.g., nitrile). Work in a chemical fume hood to minimize inhalation exposure and to protect from light.
• Light sensitivity: Protect from light (wrap containers in foil or use amber vials) to preserve fluorescence and minimize photodegradation.
• Incompatibilities: Avoid strong oxidizers or strong reducing agents that may affect the nitro group; avoid strong bases/acids if the linker is hydrolysis-sensitive (actual linkage not specified—exercise caution).
• First aid (overview; defer to SDS):
  • Inhalation: Move to fresh air; seek medical attention if symptoms persist.
  • Skin/eye contact: Rinse with water for at least 15 minutes; remove contaminated clothing; obtain medical advice.
  • Ingestion: Rinse mouth; do not induce vomiting; seek medical attention.
• Waste: Collect solutions and solids as hazardous organic waste; do not discharge to drains. Follow institutional and local regulations.

Always consult the SDS for authoritative, product-specific safety information.

Solvent Selection

Applicability: NBD-Fructose is a small-molecule fluorescent probe; solvent choice primarily impacts solubility, fluorescence intensity, and assay compatibility.

General guidance (literature, not item-specific specs)

• Likely solubility profile: Good solubility expected in DMSO, DMF, methanol; variable water solubility depending on linkage and salt form. Start with dry DMSO to prepare a concentrated stock (e.g., 1–10 mM), then dilute into aqueous buffers with 0.1–1% final DMSO where compatible.
• Polarity/photophysics: NBD emission is environment-sensitive (stronger in less polar/aprotic media; quenched in protic/wet environments). Selecting buffer and co-solvent can tune signal-to-noise.
• Buffer compatibility: Neutral buffers (PBS, HEPES) are generally suitable. Avoid strong base/acid if the linkage is hydrolysis-prone (exact linkage for this item not specified).

Small comparison (literature trends)

• DMSO: Excellent solvency; may enhance brightness pre-dilution; keep final DMSO low for cells/enzymes.
• MeOH/EtOH: Good solvency; can alter emission; avoid high percentages in live-cell assays.
• Aqueous buffers: Biologically relevant but may reduce quantum yield; add minimal organic cosolvent for solubilization.

Practical tips

• Determine λex/λem in your chosen matrix; NBD often excites near 465 nm and emits near 535 nm, but maxima shift with solvent polarity.
• Filter or centrifuge working solutions to remove particulates prior to optical measurements.

Storage and Reconstitution

Item-specific conditions (from Product Data)

• Storage: Store at −20 °C.
• Shipping: Shipped in ice chest with ice pads.

General handling and reconstitution guidance (not item-specific specs)

• Protect from light at all times; use amber vials or wrap containers in aluminum foil.
• Allow the container to equilibrate to room temperature in the dark before opening to minimize moisture condensation.
• Reconstitution: If solid, prepare a concentrated stock in anhydrous DMSO (e.g., 1–10 mM). If supplied as a solution, verify solvent and concentration on the label/CoA.
• Aliquoting: Divide stock into single-use portions to avoid repeated freeze–thaw cycles. Store aliquots at −20 °C in tightly sealed amber microtubes with desiccant if feasible.
• Working solutions: Prepare fresh in buffer immediately before use. Avoid prolonged storage of aqueous solutions; NBD fluorescence can diminish upon light and hydrolytic exposure.
• Stability: Exact shelf life and stability data are not specified for this item; refer to CoA/SDS. As a rule of thumb for NBD conjugates, minimize exposure to heat, light, and moisture.

Always refer to the product label and CoA/SDS for definitive, lot-specific instructions.

Structure and Identity

Item-specific facts (from Product Data)

• Product name: NBD-Fructose (SKU N1452088)
• CAS: 940961-04-6
• InChIKey: Not specified for this item; refer to CoA/Spec Sheet. (Listing shows “290585”, which is not a standard InChIKey format; consult CoA.)
• SMILES: 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.

General description (chemistry/literature)

• NBD-Fructose denotes a fructose molecule covalently labeled with the NBD fluorophore (7-nitrobenz-2-oxa-1,3-diazol-4-yl). NBD is a small, electron-deficient nitrobenzofurazan ring system that is widely used for fluorescent tagging of amines, thiols, and carbohydrates (literature).
• Structural features: one carbohydrate (fructose, a ketohexose) linked to an NBD aromatic heterocycle. The exact linkage position on fructose and any spacer length are not specified in this listing.
• 2D structure (verbal): a six-carbon sugar scaffold bearing multiple hydroxyl groups and a ketone-derived center, coupled via a single covalent tether to a nitro-substituted benzofurazan ring (N–O–N diazole within the benzofurazan). The NBD ring carries a nitro group para to the attachment site (literature on NBD core).

Synthetic Utility

Applicability: NBD-Fructose is typically an end-use probe rather than a versatile synthetic building block.

General synthetic/analytical contexts (literature)

• Probe in method development: Useful as a fluorescent tracer in optimizing chromatographic separations or studying carbohydrate derivatization reactions, given its strong, trackable signal.
• Derivatization/Conjugation: If the supplied NBD-fructose retains a free handle (not specified here), it might be coupled to surfaces, polymers, or biomolecules for assay development. Because the presence of any reactive group is not specified for this item, treat it as a neutral probe unless confirmed by CoA.
• Retrosynthetic perspective: NBD labels are commonly introduced via nucleophilic substitution on NBD-chloride (NBD–Cl) or coupling to aminooxy/amine-functionalized sugars (literature). This information is useful for interpreting potential degradation pathways (e.g., hydrolysis of oxime/amide/ether linkages).

Best practices

• When using NBD-fructose as an internal standard or tracer, verify spectral nonoverlap with other assay components and confirm linear fluorescence response over the working concentration range.

Target Specificity

Item-specific data

• No target/antigen specificity, clone, isotype, or species reactivity is provided for this product. It is a small-molecule probe, not an antibody or biologic. Refer to application literature and your experimental controls to define specificity in your system (e.g., competition with unlabeled fructose, use of transporter knockouts).