NBD-FTY720 phenoxy hydrochloride , CAS No.2319882-09-0

CAS: 2319882-09-0 Cat. No.: N1452061 Summenformel: C₂₃H₃₂ClN₅O₆ Molekulargewicht: 509.98

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Übersicht

NBD-FTY720 phenoxy hydrochloride is a fluorescently-labeled analog of FTY720. FTY720 is a potent sphingosine 1-phosphate (S1P) receptors modulator.

Specifications

Storage

Store at -20°C

Verschickt in

Ice chest + Ice pads

Dieses Produkt erfordert Kühlkettenversand. Grundversand und andere Economy-Optionen sind nicht verfügbar.

Namen und Kennungen

• Molekulargewicht: 509.98

Application Protocols

No manufacturer‑validated protocols are provided for this SKU. The following are general, literature‑based procedures for NBD‑labeled lipidic probes; optimize for your system.

A. Live‑cell fluorescence imaging (general)

• Prepare 10 mM DMSO stock under low light. Dilute to 1 μM in HBSS + 10 mM HEPES + 0.1% fatty‑acid‑free BSA.
• Seed cells on glass‑bottom dishes; wash with warm imaging buffer.
• Add probe solution (final DMSO ≤0.1% v/v). Incubate 15–30 min at 37°C protected from light.
• Wash 2–3× with imaging buffer. Image with 470/40 nm excitation and 525/50 nm emission filters (FITC/GFP settings). Minimize exposure.

B. Flow cytometry uptake assay (general)

• Prepare 1 mM DMSO stock. Dilute to 50–250 nM in PBS + 0.1% BSA.
• Harvest and resuspend cells at 1×10^6 cells/mL. Incubate with probe 15–20 min at 37°C or on ice for surface‑restricted binding.
• Wash twice with cold PBS + 0.1% BSA. Analyze in FITC channel. Include unstained and vehicle controls; consider unlabeled competitor for specificity.

C. Liposome partitioning (general)

• Prepare SUVs/LUVs from defined lipid mixtures. Titrate probe from 10 nM to 1 μM in buffer.
• Monitor fluorescence enhancement/shift upon addition of vesicles to estimate partitioning; correct for inner‑filter effects.

Notes

• Protect solutions from light; use amber consumables.
• Validate concentrations to avoid membrane perturbation.
• Item‑specific extinction coefficients or quantum yields are not provided; determine instrument‑specific calibration empirically.

Biological Roles

Scope note: The following summarizes general, literature‑based roles of NBD‑labeled FTY720 derivatives as research probes. No therapeutic or clinical claims are made. Item‑specific biological data are not provided in the Product Data.

General roles (literature)

• Sphingolipid‑mimetic probe: Fingolimod (FTY720) analogs mimic key features of sphingoid bases. NBD tagging provides a compact fluorophore to visualize uptake, membrane partitioning, and intracellular routing.
• Environment‑sensitive reporter: NBD fluorescence increases and blue‑shifts in nonpolar environments, enabling readouts for membrane insertion versus aqueous residence.
• Trafficking and internalization: Useful in tracking time‑dependent internalization in live cells using confocal microscopy or flow cytometry, aiding studies of lipid handling pathways.
• Assay tracer: Employed in competitive binding or transport assays to estimate affinity or transport kinetics for sphingolipid‑interacting proteins (e.g., kinases, transporters) without radioactive tracers.

Best practices (general)

• Validate that NBD tagging does not alter the biological phenomenon under study; include unlabeled analogs as controls.
• Titrate concentrations to minimize off‑target membrane perturbation; typical working ranges are low nM to low μM depending on system.
• Monitor photobleaching and potential phototoxicity under high illumination; minimize exposure times and use appropriate filters.

Item‑specific biological targets, KD values, or pathways are not specified for this product; consult primary literature and perform in‑house validation for your biological system.

Buffer Applications

Relevance: As a fluorescent probe, this compound is used in buffered media rather than serving as a buffer component.

General buffer guidance for NBD‑labeled lipidic probes (not item‑specific)

• Stock preparation: Dissolve in anhydrous DMSO (e.g., 1–10 mM). For working solutions, dilute into:
  • PBS or HBSS with 0.05–0.1% fatty‑acid‑free BSA to reduce adsorption and improve delivery.
  • Alternative carriers: 0.01–0.05% Tween‑20 or 2–5 mM methyl‑β‑cyclodextrin depending on the assay and cell tolerance.
• pH considerations: Maintain physiological pH (7.2–7.4) for live‑cell work; avoid strong base/acid which can degrade NBD or alter protonation state of amines.
• Ionic strength: Standard PBS or HEPES‑buffered saline is typically suitable; low divalent cation conditions may be preferred in some binding assays to minimize aggregation.
• Light protection: Use amber tubes/vials and minimize ambient light during dilutions.

Example working preparations (illustrative; validate for your system)

• Flow cytometry: 1 mM DMSO stock → dilute to 50–250 nM in PBS + 0.1% BSA; incubate cells 10–30 min at 37°C; wash with buffer.
• Microscopy: 1–5 μM in imaging buffer (e.g., HBSS + 10 mM HEPES + 0.1% BSA); image with blue excitation and GFP/FITC‑like emission filters; adjust exposure to limit bleaching.

Note: No item‑specific buffer recipes are provided; optimize empirically.

Green Alternatives

While the probe itself is fixed, greener choices can be made in solvent systems and workflows used with NBD‑FTY720‑type reagents.

Greener practice options (general guidance)

• Prefer aqueous buffers with minimal DMSO: Use the lowest effective DMSO co‑solvent (≤0.1–0.5% v/v) and incorporate biocompatible carriers (BSA, cyclodextrins) to limit organic solvent load.
• Replace high‑hazard solvents: If method development considered DMF, consider glycerol, PEG‑400 (small %), or ethanol as co‑solvents when compatible with biology.
• Minimize single‑use plastics: NBD probes can adsorb to plastics; consider glass vials and reusable glassware to both improve dosing accuracy and reduce waste.
• Energy use: Store at −20°C as specified, but ship and aliquot to reduce freezer door openings and freeze–thaw cycles.

Comparison of common co‑solvents (general; not item‑specific specs)

• DMSO: High solvating power, low volatility; moderate environmental impact; good choice at low %.
• Ethanol: Renewable feedstock; higher volatility; biological tolerance up to low %; may reduce solubility vs DMSO.
• 2‑MeTHF/CPME: Greener ethers but generally unnecessary for this class and less compatible with aqueous assays.

Waste considerations

• Collect DMSO/DMF solutions as hazardous waste; avoid chlorinated solvents in cleanup where possible; use aqueous detergent washes first.

These alternatives focus on process greenness; the chemical identity of the probe remains unchanged.

Pharmaceutical Uses

This product is intended strictly for research use only; not for diagnostic, therapeutic, or clinical use.

Non‑clinical, research‑stage applications (general)

• Assay development: Fluorescent tracer in high‑content imaging, flow cytometry, or plate‑based binding assays related to sphingolipid biology.
• ADME/uptake models: Qualitative visualization of cellular uptake or membrane partitioning in preclinical systems.
• Analytical reference: Fluorescence‑detected HPLC/LC‑MS method development for sphingolipid‑mimetic probes.

Formulation‑style considerations for lab use (general)

• Vehicle: DMSO stock with final assay vehicle ≤0.1–0.5% v/v in aqueous buffers to maintain cell health and signal quality.
• Excipient analogs: BSA or mild nonionic surfactants (Tween‑20) can serve as carriers to reduce nonspecific adsorption.

Regulatory/pharmacopeia

• No pharmacopeial monograph is expected for this specialized research probe.

Item‑specific excipient status, GMP grade, or clinical formulation data: Not specified for this item; refer to CoA/Spec Sheet. Always ensure use is confined to laboratory research contexts.

Physical Properties

Item‑specific physical data

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

General/literature characteristics for NBD‑labeled lipidic probes (not specifications for this item)

• Fluorescence: NBD typically shows λex ≈ 460–480 nm and λem ≈ 525–545 nm; emission is polarity‑sensitive and can blue‑shift and intensify in nonpolar environments.
• Solubility: Often readily soluble in anhydrous DMSO and DMF; limited intrinsic aqueous solubility without surfactants, cyclodextrins, or serum proteins.
• LogP: Amphiphilic; effective partitioning into membranes is common for FTY720 analogs (exact logP for this specific item not specified).
• pKa: The tertiary amine (where present) typically exhibits pKa in the 8–10 range in related analogs, favoring protonation in neutral/acidic media (literature; item‑specific value not specified).
• Physical state: Typically solid at room temperature for HCl salts of such probes (item‑specific state not specified).

Optical handling notes (general)

• NBD is moderately photostable; protect from light. Fluorescence can be quenched by high polarity solvents and by nitroaromatic photo‑bleaching pathways.

Always consult the product CoA/Spec Sheet and SDS for authoritative, item‑specific physical parameters.

Quality and Grades

Item‑specific quality information

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

How to interpret grades for fluorescent probes (general guidance)

• Research grade: Typical for imaging probes; emphasizes chemical identity and chromatographic purity. For quantitative binding studies, review residual solvent, counterion stoichiometry, and dye isomer content on the CoA.
• HPLC grade materials (when offered): Low UV background and minimized non‑fluorescent impurities; facilitates quantitative fluorescence assays and clean chromatograms.
• Salt form: Hydrochloride salts provide improved handling and reduce volatility; confirm equivalence (free base vs. HCl) when preparing molar stocks. CoA typically lists % as free base and chloride content.
• Identity confirmation: Expect 1H/13C NMR, HRMS, and HPLC purity traces. For fluorophores, spectral confirmation (λex/λem) is sometimes appended.

What to check on receipt (practical checklist)

• Match CAS (2319882‑09‑0), lot number, and net content.
• Verify purity and counterion content; note if multiple NBD positional isomers are controlled.
• Review residual solvents and water content if your assay is sensitive to these. If unspecified: Not specified for this item; refer to CoA/Spec Sheet.

Stabilizers/additives

• Any stabilizers or antioxidants are: Not specified for this item; refer to CoA/Spec Sheet.

For trace specifications (metals, water, peroxides, UV cutoff): Not specified for this item; refer to CoA/Spec Sheet.

Reaction and Applications

Focus area: As a fluorescent probe, NBD‑FTY720 phenoxy hydrochloride is primarily applied in biochemical and cell‑biology assays rather than as a synthetic reagent. No manufacturer application notes were provided for this SKU; the following are literature‑based, general uses of NBD‑labeled FTY720 analogs.

Typical applications (general; not item‑specific claims)

• Membrane association and trafficking studies: Monitor incorporation into lipid bilayers, endocytosis/exocytosis, and intracellular localization via NBD fluorescence.
• Receptor/target engagement assays: Serve as a fluorescent tracer in binding, competition, or internalization assays involving sphingolipid‑related proteins; quantify with plate readers, flow cytometry, or microscopy.
• Lipid transport and metabolism: Track uptake and redistribution in live cells or vesicles; NBD’s environment‑sensitive emission reports on partitioning states.
• Analytical reference: Use as a standard for HPLC‑FLD or LC‑MS method development for fluorescent sphingolipid analogs.

Methodological tips (general)

• Prepare concentrated anhydrous DMSO stocks (e.g., 1–10 mM); filter through 0.22 μm PTFE if particulates are observed.
• Protect from light; minimize time in bright excitation to reduce photobleaching. Use neutral density filters for long time‑lapse imaging.
• Include carrier proteins (BSA) or mild detergents (Tween‑20) to reduce nonspecific adsorption to plastics and to improve dosing accuracy at low nM–μM levels.
• Calibrate fluorescence with solvent‑matched blanks; NBD intensity varies with microenvironment polarity.

Note: Specific binding constants, kinetics, or cellular targets for this exact item are not provided; verify suitability in your assay context.

Reaction Conditions

As supplied, this product is a ready‑to‑use fluorescent probe rather than a reagent for bond‑forming reactions. Accordingly, there are no “reaction conditions” in the synthetic sense that apply to its routine use.

General operational conditions for use in assays (literature guidance; not item‑specific specifications)

• Stock solutions: 1–10 mM in anhydrous DMSO; store protected from light at −20°C. Warm to room temperature and vortex before use.
• Working concentrations: Typically 10 nM to 5 μM depending on assay sensitivity and cell type. Start with a dilution series to identify the linear range of fluorescence without perturbing membranes.
• Incubation: 10–60 minutes at 25–37°C for uptake/internalization assays; shorter times for surface binding studies on ice (to limit endocytosis).
• Detection: Excitation 460–480 nm; emission 520–540 nm using FITC/GFP‑like filter sets; adjust detector gain to avoid saturation.
• Controls: Include vehicle (DMSO) controls, unlabeled analog competitors, and temperature controls (e.g., 4°C) to estimate active versus passive uptake.

If employing chemical reactions with NBD reagents (context only)

• Reactions of NBD‑Cl/NBD‑F with amines are typically run in polar aprotic solvents with mild base (e.g., NEt3) at 0–25°C; workups under low light. This is background information and not a procedure for the current product.

Always validate conditions for your specific biological or analytical platform.

Safety and Handling

Item‑specific hazard data

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

General laboratory safety guidance for fluorescent lipidic amine salts (literature/practice)

• PPE: Use lab coat, safety glasses, and appropriate chemically resistant gloves (e.g., nitrile). Handle powders and DMSO solutions in a fume hood to avoid inhalation/dermal exposure.
• Incompatibilities: Avoid strong oxidizers, strong bases/acids for extended periods (possible degradation of NBD). Avoid reactive acylating/alkylating agents. Keep away from light and moisture to minimize hydrolysis and photodecomposition.
• Peroxide risk: Not applicable to the solid; if using ether co‑solvents, observe normal peroxide checks (solvent‑related, not probe‑specific).
• First aid (overview; defer to SDS): If on skin, wash with soap/water. If in eyes, rinse cautiously with water for several minutes and remove contact lenses. If inhaled, move to fresh air. If ingested, rinse mouth; do not induce vomiting. Seek medical attention for any exposure concerns.
• Waste: Collect solutions and contaminated disposables as hazardous organic waste. Do not discharge to drains.
• Light sensitivity: Store and handle under low light; use amber vials or foil wrap to preserve fluorescence.

Always consult the product’s SDS for authoritative hazard classification, exposure controls, and emergency procedures.

Solvent Selection

Practical solvent guidance for NBD‑FTY720‑type amphiphilic probes (general/literature; not item‑specific specifications)

• Primary stock solvents

  • DMSO (anhydrous): Preferred for 1–10 mM stocks; excellent solvating power for amphiphilic salts; low volatility; miscible with aqueous buffers upon dilution.
  • DMF (anhydrous): Alternative to DMSO when viscosity is a concern; ensure high purity to minimize background.

• Aqueous use

  • Direct water/PBS solubility can be limited; pre‑dissolve in DMSO, then dilute into buffer containing 0.05–0.1% surfactant (e.g., Tween‑20) or 0.1% fatty‑acid‑free BSA to reduce aggregation and adsorption.
  • Cyclodextrin inclusion or serum proteins can enhance apparent solubility and delivery to membranes.

• Miscibility profile (general)

  • Miscible: DMSO, DMF, alcohols (MeOH, EtOH) typically dissolve the probe; water requires co‑solvent or carriers.
  • Less suitable: Nonpolar alkanes without a polar modifier; risk of precipitation and dosing variability.

• Choosing among options

  • For live‑cell imaging: Use DMSO stock; keep final DMSO ≤0.1–0.5% v/v to preserve cell health.
  • For biochemical binding assays: Consider buffer with 0.01–0.05% detergent to minimize nonspecific adsorption to plastics.
  • For lipid vesicle work: Pre‑mix in a small volume of EtOH or DMSO before adding to lipid films or vesicle suspensions, then remove bulk solvent by gentle evaporation if needed.

Note: Exact solubility limits and solvent cutoffs are not specified for this item; consult the CoA/Spec Sheet and validate empirically.

Storage and Reconstitution

Item‑specific instructions (from Product Data)

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

General best practices for this class of probes (supplemental guidance; not item‑specific specifications)

• Light protection: Store in amber vials or wrap in foil to minimize photodegradation of NBD.
• Atmosphere: Keep tightly capped with desiccant to limit moisture ingress; avoid repeated exposure to ambient humidity.
• Aliquoting: Upon first opening, prepare small aliquots (e.g., 50–200 μL) to minimize freeze–thaw cycles and headspace.
• Reconstitution: Prepare concentrated stocks (e.g., 1–10 mM) in anhydrous DMSO. Mix thoroughly (vortex/sonicate briefly if needed) until fully dissolved. If insoluble residues persist, warm gently to room temperature and re‑vortex; avoid prolonged heating.
• Working solutions: Dilute DMSO stocks into buffer with carriers (0.05–0.1% BSA or 0.01–0.05% Tween‑20) to limit adsorption; keep final DMSO ≤0.1–0.5% v/v as assay permits.
• Stability: Short‑term handling at room temperature is acceptable during weighing and solution prep; promptly return to −20°C. Avoid >3–4 freeze–thaw cycles by using aliquots.
• Disposal: Treat unused solutions and contaminated materials as hazardous organic waste.

Any item‑specific stabilizers, exact shelf life, or solution stability: Not specified for this item; refer to CoA/Spec Sheet.

Structure and Identity

Brief overview: NBD‑FTY720 phenoxy hydrochloride is a fluorescent fingolimod (FTY720) derivative bearing an NBD (7‑nitrobenz‑2‑oxa‑1,3‑diazol‑4‑yl) fluorophore; supplied as the hydrochloride salt for research use.

Item‑specific facts (from Product Data)

• SKU: N1452061
• Product name: NBD-FTY720 phenoxy hydrochloride
• CAS: 2319882-09-0
• Storage: Store at −20°C
• Shipped: Ice chest + ice pads
• Research use: For research use only

Structure identifiers (this item)

• 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 (general/literature description of class)

• NBD fluorophore: An electron‑poor nitrobenzoxadiazole ring system that is small, environment‑sensitive, and typically excitable in the blue range.
• FTY720 core: Amphiphilic sphingolipid‑mimetic scaffold; presence of a tertiary amine often protonated as HCl salt to enhance handling and solubility in polar media.
• Phenoxy linkage: An aryl–O– moiety connecting the probe to the lipid‑like scaffold (literature, exact substitution for this item not specified).
• 2D description: One compact aromatic NBD ring fused to a 1,3,4‑oxadiazole‑like heterocycle bearing a nitro group; attached via an ether/amine spacer to a hydrophobic chain/aryl unit reminiscent of fingolimod; terminal amine protonated as hydrochloride (general depiction; consult CoA for exact regiochemistry).

Note: Exact regiochemistry, counterion stoichiometry, and stereochemical elements are not specified for this catalog item; consult the CoA/Spec Sheet for definitive identity details.

Synthetic Utility

Primary role: final fluorescent probe. It is not typically used as a synthetic intermediate.

Contextual notes (general; literature)

• NBD installation chemistry: NBD groups are commonly introduced via NBD‑Cl (4‑chloro‑7‑nitrobenzofurazan) or NBD‑F (4‑fluoro‑7‑nitrobenzofurazan) reacting with amines/thiols under mild basic conditions. While informative for understanding the probe’s origin, the current product is supplied pre‑functionalized.
• Salt form benefits: The hydrochloride salt often improves handling and enables accurate massing for stock solutions by defining the protonation state; confirm equivalence (free base vs. HCl) from CoA.
• Reference standard: May serve as a synthetic reference for chromatographic method development or as a comparator in structure–activity studies of new analogs.

If modifications are required (general strategies)

• Spacer/handle chemistry: Derivatives of FTY720 can be diversified at the amino functionality; however, NBD substitution typically occupies this handle, limiting further derivatization without deconstruction.
• Conjugation alternatives: For orthogonal tagging, synthetic routes often move the fluorophore to a side chain spacer rather than the core amine.

Item‑specific reactivity, protecting groups, or impurity profiles are not provided; consult the CoA/Spec Sheet and primary literature if planning structural analog synthesis.

Target Specificity

Item‑specific target/antigen information

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

General note

• This product is a small‑molecule fluorescent probe, not an antibody or biologic. Therefore, conventional immunoassay attributes (antigen name, epitope, clone, isotype, species reactivity) do not apply.
• In literature, NBD‑labeled FTY720 derivatives are used as tracers in assays involving sphingolipid‑related proteins; however, specific targets, KD/IC50 values, and selectivity profiles for this exact catalog item are not provided and should be determined empirically in the intended assay system.