LysoFP-NH2 , CAS No.69408-85-1

CAS: 69408-85-1 Cat. No.: L1450271 Formule: C₁₈H₁₉N₃O₃ Poids moléculaire: 325.36

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Vue d’ensemble

LysoFP-NH2 is a lysosomal-targeted fluorescent probe with amino acid functionalized derivatives. LysoFP-NH2 can be used to detect lysosomal NO levels in living cells.

Specifications

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.

Noms et identifiants

• Poids moléculaire: 325.36

Application Protocols

Item-specific (from Product Data)

• Tested applications, recommended dilutions, and positive controls: Not specified for this item; refer to CoA/Spec Sheet.

General example workflow for live-cell lysosomal staining (not item-specific; optimize empirically)

• Stock: Prepare 1–10 mM stock in anhydrous DMSO. Protect from light.
• Cell prep: Culture cells on glass-bottom dishes. Replace media with phenol red–free, pre-warmed imaging buffer.
• Working solution: Dilute stock to a trial range (e.g., 50–500 nM or 1–5 µM depending on chromophore potency). Maintain final DMSO ≤0.1–0.5%.
• Incubation: 10–30 min at 37°C, 5% CO2; protect from light.
• Wash: Rinse 2–3× with dye-free buffer; optional 10–15 min chase.
• Imaging: Use appropriate excitation/emission filters for the chromophore family. Minimize exposure to limit photobleaching/phototoxicity. Acquire z-stacks if assessing organelle distribution.
• Controls: Include cells treated with a known lysosomal marker and cells subjected to lysosomal alkalinization (e.g., brief bafilomycin A1 or NH4Cl treatment) to validate probe response.

Note: These steps are illustrative and not validated for LysoFP-NH2 specifically.

Biological Roles

Item-specific (from Product Data)

• No biological activity or target information is specified for this item; it is sold for research use only.

General background for lysosome-targeted fluorescent probes (not item-specific)

• Mechanism of accumulation: Many lysosomal probes exploit weak-base trapping. Unprotonated amines diffuse across membranes; in the acidic lysosomal lumen (pH ~4.5–5.0), they protonate and become membrane-impermeant, accumulating within lysosomes.
• Applications: Visualization of lysosomal number, size, pH heterogeneity, and trafficking; assessment of autophagy and lysosomal membrane permeabilization in cell studies. Co-staining with organelle markers helps validate specificity.
• Environmental sensitivity: Some chromophores change intensity or spectral properties with pH, viscosity, or polarity, enabling functional readouts of lysosomal state.
• Considerations: Overloading or prolonged illumination can perturb lysosomal function. Optimize dye concentration and exposure to minimize phototoxicity and avoid artifactual lysosomal alkalinization or membrane damage.

Note

• The exact probe structure, pKa, photophysics, and selectivity of LysoFP-NH2 are not provided here. For any biological inference, verify performance empirically in your system and consult the CoA/Spec Sheet if available.

Buffer Applications

Item-specific (from Product Data)

• No buffer formulation guidance is provided for this item.

General guidance for using lysosomal probes in aqueous buffers (not item-specific)

• Stock preparation: Prepare concentrated stocks in anhydrous DMSO. Dilute into pre-warmed physiological buffers (HBSS, PBS) immediately before use; typical final DMSO ≤0.1–0.5% v/v to maintain cell health.
• pH considerations: Maintain extracellular buffer near pH 7.2–7.4 to preserve normal lysosomal acidification gradients. Avoid strong amine-containing buffers (e.g., high Tris) during conjugation chemistry with activated esters, as they can quench reactions.
• Additives: 0.1% BSA can reduce nonspecific adsorption to plasticware. For live-cell imaging, phenol red–free media can lower background fluorescence.
• Washing: After loading cells, wash with dye-free buffer to reduce cytosolic/background signal; allow a short chase period if the protocol requires lysosomal accumulation equilibrium.

Not typically applicable as a buffering agent

• LysoFP-NH2 itself is not a buffer reagent. Its role, if used as a probe, is as a fluorescent reporter rather than as a component to control pH or ionic strength.

Green Alternatives

General guidance (not item-specific)

• Solvent choice has the greatest sustainability impact for dye handling. Where feasible, prefer lower-toxicity, lower-VOC, and biorenewable solvents.

Solvent alternatives for stock preparation and synthesis

• Replace DMF/NMP with:
  • Dimethyl sulfoxide (DMSO): Widely accepted for biological stocks; lower chronic toxicity profile than DMF/NMP.
  • Cyrene (dihydrolevoglucosenone) or GBL alternatives for certain couplings (method-dependent); verify dye stability.
  • 2-Methyltetrahydrofuran (2-MeTHF) or CPME for extractions and non-aqueous workups instead of chlorinated solvents; check dye solubility.
• Aqueous/biobased media: For conjugation to biomolecules, consider aqueous buffer couplings using water-soluble carbodiimides (EDC) with minimal organic co-solvent.

Process and waste minimization

• Run small-scale pre-screens to determine minimal effective dye concentration, reducing solvent usage.
• Use amber vials to extend shelf life and reduce waste from degraded stocks.
• Adopt microscale solid-phase or flow-based coupling to cut solvent volumes.

Tradeoffs

• Greener solvents may alter reaction rates or solubility; ensure pilot testing. Some biobased solvents can react with activating agents or degrade sensitive chromophores—evaluate compatibility before scale-up.

Pharmaceutical Uses

Item-specific (from Product Data)

• No pharmacopeial designation or formulation role is specified. For research use only.

General considerations (not item-specific; no therapeutic claims)

• Process analytics and R&D: Fluorescent probes can be used in pharmaceutical research workflows for tracking subcellular localization, evaluating lysosomotropic properties of candidate compounds, and visualizing intracellular delivery of formulations in cell-based assays.
• Excipient compatibility: If incorporated into nanoparticle or liposomal test articles as a tracer, ensure the dye’s amine does not undesirably react with activated excipients or crosslinkers. Validate leaching and stability under storage conditions.
• Regulatory perspective: Research-use-only dyes are not intended for human administration. Any use in GMP-enabling studies would require rigorous qualification and documentation not provided here.

Documentation

• For impurity profiles, residual solvents, and stability data suitable for internal quality systems, request the CoA/Spec Sheet for this SKU.

Physical Properties

Item-specific (from Product Data)

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

Typical/literature expectations for amine-terminated fluorescent probes (not item-specific)

• Physical state: Often supplied as a solid (powder) or film; hygroscopicity and photolability vary by chromophore.
• Solubility: Many fluorophores dissolve well in polar aprotic solvents (DMSO, DMF, NMP). Aqueous solubility depends on ionizable groups/salts; primary amines can form water-soluble hydrochloride salts.
• Stability: Chromophores may be light-sensitive; storage under inert atmosphere and minimal headspace moisture is often beneficial.
• Acid–base: If a primary amine is present, conjugate acid pKa commonly ~8–10 (literature, scaffold-dependent), influencing lysosomal trapping in acidic pH (~4.5–5.0).
• Partitioning: logP/logD varies widely with scaffold and ionization state; no item-specific value is available.
• Spectroscopy: Absorption/emission maxima are chromophore-dependent. Not specified for this item; refer to CoA/Spec Sheet if provided.

Do not treat these literature expectations as specifications. For precise values (mp/bp, density, solubility, refractive index, UV/Vis maxima, extinction coefficient), consult the CoA/Spec Sheet for this SKU.

Quality and Grades

Item-specific (from Product Data)

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

General guidance (not item-specific)

• Research-grade fluorescent probes are typically supplied at high chemical purity to minimize background fluorescence and side reactions during conjugation. Purity is commonly verified by HPLC/UPLC with UV/Vis detection and, in many cases, MS.
• If provided, “HPLC grade (dye)” or similar annotations indicate low levels of fluorescent impurities and residual reagents; this is distinct from solvent HPLC grade. For conjugation-ready amines, residual protecting groups and inorganic salts should be minimal.
• Spectral QC: Some suppliers include absorbance/emission maxima and extinction coefficients. These are useful for instrument setup and quantitation. If not provided here, verify on your instrument prior to critical experiments.
• Stabilizers: Some dyes are supplied with small amounts of stabilizers or as TFA/HCl salts to improve shelf life and handling. Any stabilizer or counter-ion used should be disclosed on the CoA/Spec Sheet.
• Lot-to-lot documentation: For sensitive imaging applications, retain the CoA and consider performing a small pilot labeling run to confirm performance before scaling.

Reaction and Applications

Item-specific (from Product Data)

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

General/literature applications for an amine-terminated lysosome-oriented fluorescent probe (not item-specific)

• Live-cell imaging: Weakly basic, amine-bearing fluorophores often accumulate in acidic organelles (lysosomes). When appropriately structured, these dyes can enable visualization of lysosomal morphology, trafficking, and autophagic flux.
• Bioconjugation: The terminal –NH2 can be coupled to carboxylates (EDC/HOBt, HATU, PyBOP) to generate amides, or reacted with NHS esters, isothiocyanates (to form thioureas), or activated carbonates (to form carbamates). This enables tethering to ligands, peptides, nanoparticles, or surfaces while preserving targeting.
• Sensor construction: The amine handle can be used to append environment-responsive motifs (pH, enzymes), yielding ratiometric or turn-on lysosomal probes.
• Materials and diagnostics R&D: Incorporation into polymers, liposomes, or beads to create lysosome-tracking carriers for in vitro studies.

Practical tips (general)

• Protect from light and moisture during synthesis and handling. Use anhydrous solvents and inert atmosphere for coupling steps to minimize hydrolysis/oxidation.
• Verify excitation/emission on your instrument; lysosomal dyes often excite in the blue–green range, but this is chromophore-dependent. Determine bleed-through and set suitable filters to avoid crosstalk with other channels.
• Validate localization with a counterstain (e.g., a well-characterized lysosomal marker) to confirm targeting in your cell model.

Reaction Conditions

General conditions for common transformations of amine-terminated dyes (not item-specific; optimize per system)

• Amide coupling (carboxylic acid partner):
  • Solvent: Anhydrous DMF, DMSO, or CH2Cl2 (solubility-dependent).
  • Reagents: HATU or PyBOP (1.1–1.5 eq), base DIPEA (2–4 eq), 0–25°C, 1–16 h. For aqueous-compatible protocols, EDC (1.2–2 eq) + NHS (1.2–2 eq) in MES buffer pH 5.0–6.0, 0–25°C.
  • Notes: Minimize light exposure; monitor by LC–MS or HPLC. Avoid excess base if chromophore is base-sensitive.
• Carbamate formation (activated carbonate):
  • Solvent: Dry DMF or CH2Cl2.
  • Reagents: p-NPC or DSC-activated carbonates, base (DIPEA), 0–25°C.
• Urea/thiourea formation:
  • React with (thio)isocyanates in dry solvent; mild base; rt to 50°C.
• Reductive amination:
  • Solvent: MeOH/DMF or MeCN/DMF mixtures.
  • Conditions: Aldehyde (1.2–2 eq), NaBH3CN or picoline borane, AcOH (cat.), rt to 40°C, 2–16 h.

Workup and purification

• Quench carefully; dyes can streak on silica. Use reverse-phase HPLC (C18) with volatile buffers (e.g., 0.1% TFA or formic acid) for analytical and preparative purification.

Performance checks

• Record UV/Vis spectra before/after reaction to assess chromophore integrity. Confirm mass by ESI-MS. Determine labeling degree when conjugating to biomolecules by spectrophotometry.

These are literature-style guidelines; no item-specific validated protocol is provided.

Safety and Handling

Item-specific (from Product Data)

• Storage conditions: Store at -20°C.
• Shipping: Ice chest + ice pads.
• GHS/Signal word/H-statements/Pictograms: Not specified for this item; refer to SDS.

General safety guidance (not item-specific; defer to SDS for authoritative instructions)

• PPE: Wear lab coat, suitable gloves (e.g., nitrile), and safety glasses/goggles. Handle powders/solid dyes in a fume hood to avoid dust inhalation and to minimize exposure.
• Light sensitivity: Many fluorophores degrade upon prolonged light exposure. Minimize ambient light; use amber vials/foil wrapping.
• Incompatibilities: Strong oxidizers and strong acids/bases may degrade organic chromophores or amine functionalities. Avoid moisture if the compound is moisture-sensitive or provided as an activated salt/derivative.
• Hygiene: Avoid inhalation, ingestion, or skin/eye contact. Wash thoroughly after handling. Do not pipette by mouth. Avoid aerosol formation when preparing stock solutions.
• First aid (general): In case of skin contact, wash with soap/water. Eye contact: rinse with water for several minutes, remove contact lenses if present and easy to do; seek medical attention. Inhalation: move to fresh air; seek medical attention if symptoms persist. Ingestion: rinse mouth; seek medical attention.
• Waste: Dispose of according to institutional and local regulations for organic laboratory chemicals and dyes. Segregate halogenated vs non-halogenated solvent waste if applicable.

Research use only: This material is for laboratory research use; not for diagnostic, therapeutic, or human/animal consumption.

Solvent Selection

Item-specific (from Product Data)

• No item-specific solubility or solvent recommendations are provided. Refer to CoA/Spec Sheet.

General considerations for amine-terminated fluorescent probes (not item-specific)

• Stock solutions: Anhydrous DMSO is a common choice due to broad solvency and compatibility with biological assays at low final percentages. DMF or NMP can also be used for synthetic steps (e.g., amide couplings).
• Aqueous work: If the amine is supplied as a salt (e.g., HCl), water or aqueous buffer may be suitable; however, many hydrophobic chromophores require co-solvent (DMSO, ethanol) or surfactant assistance.
• Polarity/miscibility: DMSO and DMF are fully miscible with water, enabling convenient dilution into buffers; ensure final DMSO v/v remains compatible with cells or enzymes (commonly ≤0.1–1%).
• Degassing/dryness: For moisture-sensitive transformations (e.g., carbodiimide couplings), dry, oxygen-poor conditions reduce side reactions and dye degradation.

Comparison (general)

• DMSO vs DMF: DMSO is lower toxicity and commonly preferred for biological stock solutions; DMF may be favored for certain couplings but has higher regulatory concern. Ethanol can be a benign diluent for some assays but has limited solvency for bulky dyes.

Always confirm solvent compatibility with your assay/biology and run a small solubility test before preparing concentrated stocks.

Storage and Reconstitution

Item-specific (from Product Data)

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

General best practices for fluorescent probes (not item-specific)

• Light/moisture protection: Store in amber, airtight containers with desiccant. Consider inert gas backfill for long-term storage, especially for moisture- or oxygen-sensitive dyes.
• Aliquoting: Upon first opening, divide into single-use aliquots to minimize freeze–thaw and headspace humidity. Keep records of open dates and aliquot IDs.
• Reconstitution: If compatible, dissolve in anhydrous DMSO to prepare a concentrated stock (e.g., 1–10 mM). Filter through 0.2 µm PTFE if particulate is present. Avoid aqueous buffers for long-term stock unless the item is explicitly supplied as a water-stable salt.
• Stability: Many dye stocks are stable for months at –20°C when protected from light and moisture. Verify stability periodically by HPLC/LC–MS and absorbance/emission scans.
• Handling at bench: Thaw only the required aliquot immediately before use. Keep solutions on ice and shielded from light during setup.

For any item-specific guidance on solvent choice, concentration limits, or shelf life, consult the CoA/Spec Sheet for this SKU.

Structure and Identity

Item-specific (from Product Data)

• Product name: LysoFP-NH2 (SKU: L1450271)
• CAS: 69408-85-1
• 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.

General/literature notes (context; not item-specific)

• The designation “LysoFP-NH2” suggests an amine-terminated fluorescent probe scaffold potentially optimized for lysosomal localization. Many lysosome-targeted dyes incorporate weakly basic amines that become protonated in acidic vesicles, leading to accumulation.
• An “-NH2” suffix typically indicates a primary amine handle suitable for bioconjugation (e.g., amide formation with activated carboxylic acids) or surface attachment.

2D structural description (general expectations)

• Without an item-specific structure, functional groups and ring systems cannot be confirmed. Fluorescent probes commonly contain polyaromatic or heteroaromatic chromophores (xanthenes, coumarins, BODIPYs, cyanines, etc.) appended with targeting/solubilizing substituents. If present, a primary amine would be sp3- or sp2-linked to the chromophore via an alkyl/aryl tether.

Important

• For definitive identifiers (formula, exact structure, stereochemistry), consult the Certificate of Analysis (CoA) or Specification Sheet for this exact lot.

Synthetic Utility

General reactivity of an –NH2-terminated fluorescent probe (not item-specific)

• Amide formation: Couple with carboxylic acids or active esters to append ligands, spacers, or polymers. Reagents: EDC/HOBt, HATU, PyBOP, DIC/DMAP (conditions substrate-dependent).
• Urea/thiourea formation: React with isocyanates/isothiocyanates to tune polarity, H-bonding, and binding interactions.
• Carbamate formation: Reaction with chloroformates or activated carbonates introduces cleavable linkers or protection.
• Reductive amination: Conjugate to aldehydes/ketones (NaBH3CN, picoline borane) to build C–N linkages onto carbohydrates or oxidized polymers.
• Click-adjacent strategies: Convert the amine to azide/alkyne-bearing linkers (e.g., via NHS–PEG–N3) to enable SPAAC/CuAAC while retaining the dye core.
• Solid-phase attachments: The –NH2 group can anchor to resins or surfaces for sensor arrays and imaging plates.

Retrosynthetic value

• The amine handle provides a convergence point for late-stage diversification—useful when one wishes to graft the same chromophore onto multiple targeting motifs without resynthesizing the dye core.

Notes

• Protect the dye chromophore during harsh steps (acidic or oxidative). Confirm that coupling auxiliaries (e.g., HOBt) and bases do not quench fluorescence or cause side reactions with the chromophore.

Target Specificity

Item-specific (from Product Data)

• Target/antigen/epitope: Not specified for this item.
• Species reactivity/clone/isotype: Not applicable; no antibody or biologic details provided.

General note

• For fluorescent probes marketed for lysosomal imaging, “specificity” typically refers to preferential accumulation in lysosomes due to physicochemical properties rather than binding to a discrete protein target. Confirm localization in your system using orthogonal markers.