LysoFP-NO2 , CAS No.69408-75-9

CAS: 69408-75-9 Cat. No.: L1450375 PubChem CID: 12680935

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Overview

LysoFP-NO2 is a turn-on fluorescent probe for carbon monoxide (CO) that localizes to the lysosome. In the presence of lysosomal CO, lysoFP-NO2 is transformed into lysoFP-NH2, which is highly fluorescent. LysoFP-NO2 is selective for CO over various reactive nitrogen, oxygen, and sulfur species. It displays excitation/emission maxima of 440/528 nm, respectively, and is not cytotoxic to HepG2 cells for up to five hours when used at a concentration of 30 μM.

Specifications

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

This product requires cold chain shipping. Ground and other economy services are not available.

Names and Identifiers

• Canonical Smiles: C1COCCN1CCN2C(=O)C3=CC=CC4=CC(=CC(=C43)C2=O)[N+](=O)[O-]
• IUPAC Name: 2-(2-morpholin-4-ylethyl)-5-nitrobenzo[de]isoquinoline-1,3-dione
• InChIKey: MQBIIGAYEJQXCC-UHFFFAOYSA-N
• INCHI: 1S/C18H17N3O5/c22-17-14-3-1-2-12-10-13(21(24)25)11-15(16(12)14)18(23)20(17)5-4-19-6-8-26-9-7-19/h1-3,10-11H,4-9H2
• Isomeric SMILES: C1COCCN1CCN2C(=O)C3=CC=CC4=CC(=CC(=C43)C2=O)[N+](=O)[O-]
• PubChem CID: 12680935

Application Protocols

The following is general, literature-based guidance for lysosomal fluorescent probes; optimize for your system. Item-specific parameters (spectra, extinction coefficient, optimal dose) are not provided here and should be confirmed from the CoA/technical sheet.

Live-cell lysosome staining (general):

• Stock: Prepare 1–10 mM solution in anhydrous DMSO. Store in aliquots at −20°C, protected from light.
• Cells: Culture adherent or suspension cells to 60–80% confluence (adherent) or log phase (suspension). Pre-warm buffer or medium to 37°C.
• Working solution: Dilute stock into complete medium or HBSS/PBS to 50–500 nM final concentration. Maintain final DMSO ≤0.1–0.5% v/v.
• Incubation: 10–30 min at 37°C, 5% CO2. Gentle rocking improves uniformity. Optionally wash 1–3× with warm buffer to reduce background.
• Imaging: Acquire with filters/lasers matched to the probe. Minimize exposure to reduce photobleaching; use identical settings for comparative experiments.
• Controls: Include vehicle-only, no-dye, and colocalization with a validated lysosome marker to confirm targeting. Consider bafilomycin A1 treatment to test pH dependence.

Fixed-cell option (general):

• Some lysosome probes are compatible with fixation. If required, stain live, then fix with 3–4% paraformaldehyde (compatibility must be validated for this specific probe). Mount in antifade medium.

Data quality tips:

• Titrate concentration and incubation time for each cell type. Record background and flat-field corrections. Document lot number and preparation details for reproducibility.

Biological Roles

Context for use (general literature; not item-specific):

• Lysosomal function: Lysosomes are acidic organelles mediating macromolecule degradation, recycling, and signaling. Their pH (~4–5) and enzyme content are critical for autophagy and endocytic processing.
• Probe concept: Lysosome-targeted fluorescent probes typically exploit weakly basic moieties that accumulate via pH-driven protonation (“ion trapping”), or via ligands that recognize lysosome-associated biomolecules. Some scaffolds may be pH-sensitive, reporting acidity through intensity or spectral shifts.
• Applications in cell biology: Monitoring lysosomal abundance, morphology, motility, fusion/fission dynamics, and responses to pharmacological perturbations (e.g., V-ATPase inhibitors that alkalinize lysosomes) underpins studies of trafficking and catabolic flux.
• Colocalization: Validation commonly involves co-staining with established lysosomal markers (e.g., LAMP1 immunostaining in fixed cells or commercial lysosome trackers in live cells) and computing Pearson’s/Spearman’s correlation coefficients.

Best practices:

• Optimize probe dose and incubation to prevent lysosomal swelling or off-target accumulation in other acidic vesicles.
• Control for phototoxicity and photobleaching during time-lapse imaging; minimize light exposure and use appropriate neutral density filters.
• Consider cellular context: Lysosomal number and acidity vary with cell type and metabolic state; standardize culture conditions for comparability.

Important note: This product is for research use only. No diagnostic or clinical use is intended or implied.

Buffer Applications

This item is a small-molecule fluorescent probe rather than a buffering agent. As such, it is not typically employed to establish or maintain pH in solution.

Practical notes (general for probe use in buffered media):

• Working solutions are commonly prepared by diluting a DMSO stock into physiological buffers (e.g., HBSS, PBS) or complete culture medium immediately prior to use. Maintain DMSO at low final percentages (≤0.1–0.5% v/v) to limit solvent effects on cells.
• Buffer pH can influence lysosomal accumulation and fluorescence response. Maintain extracellular pH at ~7.2–7.4 for live-cell assays unless your experimental design requires deviation.
• Avoid high concentrations of primary amines (e.g., Tris buffer) if the probe is amine-reactive; verify compatibility for your specific probe chemistry by consulting the CoA/technical sheet.

Green Alternatives

While this product itself is a probe rather than a bulk solvent or reagent, greener practices can be applied to its handling and use. The following considerations are general (not item-specific):

• Solvent choice: Prefer DMSO over DMF for stock solutions when compatible—DMSO has a more favorable EHS profile. Use minimal organic cosolvent in working solutions (≤0.1–0.5% v/v in biological assays) to reduce environmental and biological burden.
• Waste minimization: Prepare the smallest practical aliquots to avoid disposal of expired/contaminated solutions. Consolidate dye-containing waste streams for proper disposal.
• Energy efficiency: Store as recommended (−20°C) but avoid excessive ultra-low freezers if not necessary; group shipments to reduce cold-chain footprint.
• Light management: Use amber vials to extend shelf life, reducing waste from photodegradation.

Illustrative comparison (general pros/cons for dye stock solvents):

• DMSO: Lower toxicity than DMF; biodegradable tendencies; hygroscopic and can oxidize contaminants—use fresh, high-purity grades.
• DMF: Strong solvency but reproductive-toxicity concerns and regulatory scrutiny; avoid when feasible.
• MeCN: Volatile and flammable; simplify removal but increases VOC emissions—use sparingly.

Green lab tips:

• Adopt microscale imaging workflows to reduce reagent consumption.
• Validate minimal effective staining concentrations to lower chemical use.
• Use reusable, low-autofluorescence labware to minimize plastic waste without compromising data quality.

Pharmaceutical Uses

No pharmacopeial or excipient status is provided for this item.

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

General considerations (research/manufacturing context; not item-specific):

• Fluorescent probes like lysosome-targeting dyes are used in discovery workflows (e.g., high-content screening, cellular mechanistic assays) to profile compound effects on endolysosomal pathways.
• These materials are research reagents and are not intended for human or veterinary use in drug products. There are no claims here regarding compliance with USP/EP/JP monographs.
• For regulated environments, users should qualify the reagent under their internal quality systems, documenting identity, purity, stability, and performance characteristics relevant to the intended analytical method.

Reminder: This product is for research use only; no clinical or diagnostic application is intended or implied.

Physical Properties

Item-specific physicochemical properties are not provided in this listing.

• 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: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular formula: Not specified for this item; refer to CoA/Spec Sheet.
• Melting point: Not specified for this item; refer to CoA/Spec Sheet.
• Boiling point: Not applicable for a dye-like solid; Not specified for this item; refer to CoA/Spec Sheet.
• Density: Not specified for this item; refer to CoA/Spec Sheet.
• LogP, pKa, refractive index: Not specified for this item; refer to CoA/Spec Sheet.
• UV-Vis/fluorescence maxima: Not specified for this item; refer to CoA/Spec Sheet.
• Solubility: Not specified for this item; refer to CoA/Spec Sheet.

General guidance (literature, not item-specific):

• Many lysosome-directed fluorescent probes are supplied as hydrophobic aromatic solids, often soluble in polar aprotic organics (e.g., DMSO, DMF) and sparingly soluble in water unless formulated.
• Stock solutions for cell imaging are commonly prepared at 1–10 mM in anhydrous DMSO and diluted to nanomolar–low micromolar working concentrations in aqueous buffers or culture media.
• Fluorescent properties can be pH-sensitive; lysosomal probes may display brightness changes or color shifts under acidic conditions. Exact excitation/emission values must be confirmed from the product’s CoA/technical sheet.

Quality & Grades

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

Interpretation and implications (general guidance for fluorescent probes; not item-specific):

• For imaging reagents, high chemical purity and low levels of fluorescent/non-fluorescent impurities are important to minimize background and ensure reproducible brightness. HPLC traces and spectroscopic identity (1H/13C NMR, MS, and purity by HPLC/UPLC) are typical quality indicators.
• Absence of stabilizers vs presence: Some dyes are stabilized with trace antioxidants or acid scavengers. If a stabilizer is present, it should be disclosed on the CoA; stabilizers can influence solubility and spectral baselines. This listing does not indicate any stabilizer for this item.
• Photophysical QC: For fluorescent probes, specification sheets often include excitation/emission maxima and extinction/quantum yield benchmarks. Because these are not provided here, users should confirm performance under their instrument settings.
• End-use suitability: For microscopy/flow cytometry, low particulates and minimal free acid/base are advantageous. If the grade is intended for bioimaging, expect tight controls on residual solvents and inorganic salts; verify on the CoA.

Recommendations:

• Prior to critical experiments, record a reference spectrum and run a small pilot titration to verify brightness and localization.
• Maintain lot-specific documentation (CoA, SDS) with your data to ensure traceability and reproducibility.

Reaction & Applications

This product is positioned as a life-science imaging probe rather than a synthetic reagent. Consequently, classical synthetic “reaction applications” (e.g., coupling, Grignard) are not the primary use-case.

Research applications (general/literature; not item-specific):

• Live-cell lysosomal imaging: Assessment of lysosomal integrity, morphology, and dynamics in cell culture by widefield, confocal, or high-content microscopy.
• pH-responsive readouts: Some lysosome probes display signal changes with pH; can be used in colocalization with LysoTracker-type stains to validate targeting (confirm spectral compatibility first).
• Organelle trafficking assays: Track autophagic flux or endolysosomal maturation when combined with markers for endosomes/autophagosomes, subject to appropriate assay design.

Practical considerations:

• Photophysics: Determine excitation/emission compatibility with your filter sets or lasers; perform spectral unmixing if multiplexing.
• Controls: Include vehicle (DMSO) control, non-stained cells, and colocalization with a validated lysosome marker when establishing protocols.
• Probe concentration and time course profoundly affect signal-to-noise and localization fidelity; perform titration and incubation-time optimization.

Note: Without item-specific spectral and chemical data, the above guidance is general and intended to help plan pilot experiments. For quantitative biophysical/biochemical measurements, consult the item’s technical sheet for definitive parameters.

Reaction Conditions

Not typically applicable: This product is used as an imaging probe rather than a reagent for chemical transformations.

General handling conditions for preparing working solutions (literature; not item-specific):

• Stock preparation: Dissolve in anhydrous DMSO at 1–10 mM under low light. Vortex gently to fully dissolve. Filter (0.22 µm PTFE) if particulate persists.
• Working dilution: Add stock to pre-warmed buffer or culture medium to achieve 50–500 nM (typical screening range). Keep final DMSO ≤0.5% v/v. Incubate 10–30 min at 37°C, 5% CO2 for live-cell assays, then wash if protocol requires.
• Imaging: Use excitation/emission settings matching the probe’s spectrum (consult technical sheet). Minimize exposure to limit photobleaching/phototoxicity.

Quality controls:

• Include vehicle controls and verify lysosomal localization by colocalization with a reference dye or LAMP1 antibody (fixed cells). Optimize concentration/time for each cell line.

Note: Exact concentrations, incubation times, and spectral settings must be confirmed for this specific product once item-specific photophysical data are consulted.

Safety & Handling

Safety data specific to this item are not supplied in the listing and must be obtained from the SDS.

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

General laboratory safety (good practice; not item-specific):

• Handle in a fume hood, minimize inhalation/ingestion/skin contact. Wear appropriate PPE: lab coat, safety glasses, and chemical-resistant gloves (e.g., nitrile). Avoid generating dust/aerosols.
• Many fluorescent dyes are light-sensitive; minimize ambient light exposure and store in amber containers. Prevent prolonged contact with oxidants, strong acids/bases unless intended by protocol.
• First aid (overview): In case of skin contact, wash with soap and water. Eye contact: rinse cautiously with water for several minutes; remove contact lenses if present. Ingestion/inhalation: seek medical attention; move to fresh air if inhaled. Always follow your institutional EHS procedures.
• Spill/cleanup: Absorb small spills with inert material (e.g., vermiculite), place in chemical waste. Decontaminate surfaces with suitable solvent/detergent while observing compatibility.
• Waste disposal: Collect in appropriate organic waste stream per local regulations; dyes can be persistent—avoid drain disposal.

Storage and shipping (item-specific):

• Storage: Store at −20°C (as provided). Protect from light and moisture; keep container tightly closed.
• Shipping: Ice chest + ice pads (as provided). Upon receipt, return to recommended storage.

Solvent Selection

Item-specific solubility is not disclosed. The following are general best practices for small-molecule fluorescent probes (not item-specific):

• Primary stock solvent: Anhydrous DMSO is commonly preferred due to high solvating power for aromatic dyes, low volatility, and compatibility with aqueous dilution at low final percentages (≤0.1–0.5% v/v typical in cell assays). DMF or MeCN are alternatives when DMSO is unsuitable.
• Aqueous dilution: Dilute stocks into buffered saline (e.g., HBSS, PBS) or complete culture medium immediately before use. Pre-warm and mix gently to avoid precipitation; include 0.02–0.1% serum or carrier protein if aggregation is observed (assay-dependent).
• pH considerations: Lysosomal probes are evaluated at physiological pH extracellularly but accumulate in acidic compartments; avoid strongly basic aqueous vehicles that could neutralize protonatable groups and alter partitioning.
• Avoid: High percentages of organic cosolvent in live-cell assays (generally keep DMSO ≤0.5% v/v). Avoid nonpolar hydrocarbons that lack miscibility with aqueous media.

Comparison (general):

• DMSO: Excellent solvency for dyes; biocompatible at low %; hygroscopic.
• DMF: Similar solvency; higher toxicity; more regulated in some settings.
• MeCN: Volatile, lower viscosity; moderate solvency for some dyes.

Practical tips:

• Filter 0.22 µm if particulate forms on dilution (compatibility permitting).
• Prepare single-use aliquots to prevent repeated freeze–thaw and moisture uptake.
• Validate solvent choice by examining background fluorescence and cell viability under assay conditions.

Storage & Reconstitution

Item-specific storage and shipping:

• Storage conditions: Store at −20°C (as provided). Keep tightly closed, desiccated, and protected from light.
• Shipped in: Ice chest + ice pads (as provided). Upon receipt, inspect packaging, then transfer immediately to −20°C storage.

Reconstitution and handling (general guidance; not item-specific):

• Solvent: Dissolve in anhydrous DMSO to prepare a concentrated stock (e.g., 1–10 mM). If the product is moisture-sensitive or light-sensitive, open vials briefly under low light and dry atmosphere.
• Aliquoting: Prepare single-use aliquots in amber vials or light-protective tubes to avoid repeated freeze–thaw and photodegradation.
• Working solutions: Dilute into pre-warmed buffer or culture medium immediately before use. Discard unused working solutions; do not refreeze diluted aqueous mixtures.
• Freeze–thaw: Minimize cycles. If precipitation occurs after thawing, warm gently to room temperature and vortex to re-dissolve; avoid prolonged heating.
• Stability: Specific shelf-life, stability in solution, and light sensitivity are not specified for this item; refer to CoA/Spec Sheet and SDS for definitive guidance.

Documentation:

• Record preparation date, solvent lot, and concentration on each aliquot. Maintain CoA and SDS with your lab notebook for traceability.

Research use note: For research use only.

Structure & Identity

Brief overview: LysoFP-NO2 is presented as a lysosome-directed fluorescent probe for life-science research. Item-specific structural identifiers are largely not disclosed in this listing.

• Product name: LysoFP-NO2 (research-use fluorescent probe)
• CAS: 69408-75-9 (as provided)
• PubChem CID: 12680935 (as provided)
• InChIKey: 225626 (as provided; note: this appears non-standard/incomplete; refer to CoA/SDS for the authoritative identifier)
• 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.

Structural features (general/literature context, not item-specific):

• Lysosome-targeted probes frequently incorporate weakly basic motifs (e.g., tertiary amines) that undergo proton trapping in acidic organelles (pH ~4–5), and an aromatic fluorophore scaffold for excitation/emission readout.
• The “-NO2” suffix often denotes a nitro-substituted chromophore, which can modulate electron density and photophysics (e.g., internal charge transfer, quenching/PET effects), but the exact substitution pattern for this item is not disclosed here.

2D description (general):

• Typical lysosomal probes comprise a conjugated polycyclic ring system (fluorophore core) bearing substituents that tune photophysical properties and direct subcellular localization. Without an item-specific structure, no definitive ring/functional-group assignment can be made from the current listing. Consult the Certificate of Analysis (CoA) or technical datasheet for definitive structural information.

Synthetic Utility

Primary use-case: bioimaging probe. It is not typically employed as a synthetic building block or reagent in preparative organic chemistry.

General notes (literature; not item-specific):

• Fluorophore scaffolds sometimes serve as reporters in sensor constructs, where conjugation to targeting ligands or biopolymers can be carried out via functional handles (e.g., NHS esters, maleimides, azides/alkynes). However, the functionalization state of this item is not specified here.
• If a nitro group is present in the scaffold (suggested by “-NO2” in the name), classical transformations (e.g., reduction to anilines, nucleophilic aromatic substitution on activated rings) are possible in principle, but such modifications would alter the probe’s intended photophysics and targeting and are not the product’s purpose.

Conclusion: Treat this compound as a finished imaging reagent. For custom derivatization or conjugation chemistry, consult the supplier regarding available functionalized analogs and confirm the exact structure and reactive handles on the CoA/technical datasheet.

Target Specificity

No antigen/epitope information applies; this is not an antibody or protein binder.

• Targeting concept (general; not item-specific): Lysosome-directed small molecules often accumulate via pH-dependent protonation and retention, thus showing functional specificity for acidic organelles rather than molecular selectivity for a single protein target.

Item-specific targeting parameters (e.g., Kd, selectivity indices) are not provided in this listing.