SPiDER-βGal-1 - ≥99% , CAS No.1824699-57-1

CAS: 1824699-57-1 Cat. No.: S1455228 Formula: C₃₁H₃₄FNO₈ Peso molecolare: 567.60

GRADE & PURITY ≥99%

Storage

Protected from light,Store at -20°C

Shipped In

Ice chest + Ice pads

Panoramica

SPiDER-βGal-1 is a potent probe for β-galactosidase activity and suitable for labeling live cells in culture.

Specifications

Specifiche e purezza

≥99%

Condizioni di conservazione di stoccaggio

Protected from light,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.

Purezza

≥99%

Nomi e identificatori

Peso molecolare

567.60

Application Protocols

Item-specific, validated protocols for SPiDER-βGal-1 are not provided in the Product Data. Please refer to the CoA/Spec Sheet for any manufacturer-recommended procedures.

In keeping with catalog policy for this section, we do not present generalized or literature protocols as product-validated instructions. Users should develop or adapt protocols based on their model system (e.g., lacZ reporter assays, senescence-associated β-gal imaging, lysate-based enzyme assays), performing pilot studies to optimize probe concentration, incubation time, and buffer conditions.

For method development:

• Verify instrument settings and spectral filters with an activated reference sample prepared in-house.
• Include appropriate positive and negative controls to confirm β-gal–dependent activation.
• Document all handling under low-light conditions and maintain consistent DMSO percentages across conditions.

Contact Aladdin Scientific support with your SKU and lot number for any lot-specific protocol notes that may be available.

Biological Roles

Scope: SPiDER-βGal-1 is intended for research use as a fluorogenic reporter of β-galactosidase activity. No therapeutic or diagnostic claims are made.

Biological context (general, literature):

• β-Galactosidase enzymes hydrolyze terminal β-D-galactose residues from glycoconjugates. They are widely used as lacZ reporters in molecular biology and exhibit context-dependent activity in mammalian cells (e.g., lysosomal β-galactosidase).
• Senescence-associated β-galactosidase (SA-β-gal) activity, commonly assayed near pH ~6, is a widely adopted research marker for cellular senescence in vitro. Fluorogenic β-gal probes enable sensitive detection of these activities.
• Mechanistic basis of SPiDER-type probes: The β-gal–mediated removal of a masking galactoside unmasks a latent phenolic/arylic site that undergoes an intramolecular reaction to generate a cationic fluorophore with enhanced cellular retention, supporting wash-minimized imaging workflows.

Use cases (research):

• Verification of lacZ gene expression in cells and model organisms.
• Monitoring β-gal activity changes in response to perturbations (e.g., gene knockdown, small-molecule modulators) in cell culture.
• Multiplex imaging with counterstains (nuclei, membranes) using non-overlapping spectral channels; exact channel selection depends on the specific SPiDER-βGal variant.

Caveats:

• Endogenous β-gal activities vary with cell type, pH, and subcellular localization. Include controls (pH-matched no-enzyme, competitive inhibitors) to confirm specificity.
• Validate instrument settings and filters with standards; do not assume spectral properties for SPiDER-βGal-1 without consulting the CoA/Spec Sheet.

Buffer Applications

Item-specific buffer recommendations: Not specified for this item; refer to CoA/Spec Sheet.

General buffer guidance for β-galactosidase assays (literature):

• pH selection by biological context:
  • lacZ reporter (E. coli β-gal): near-neutral buffers (phosphate or HEPES, pH ~7.0–7.4) typically yield robust activity.
  • Lysosomal β-gal: acetate or citrate buffers at acidic pH (~4–5) are common for enzymatic extracts.
  • SA-β-gal (senescence-associated): phosphate/citrate or phosphate buffers near pH ~6.0–6.5 are frequently used in cell-based assays.
• Ionic strength: Moderate salt (e.g., 100–150 mM NaCl) maintains osmolarity and can stabilize enzyme conformation; adjust empirically.
• Additives:
  • Detergents (e.g., 0.01–0.1% Triton X-100 or saponin) for fixed-cell or lysate assays to improve probe accessibility; avoid in live-cell imaging unless validated.
  • Mg2+/Ca2+: typically not required for β-galactosidase but confirm with your enzyme source.
• Cosolvent: Use minimal DMSO to deliver the probe (≤0.1–0.5% v/v final); equilibrate temperature and pH before adding the probe.

Practical notes:

• Filter-sterilize working buffers (0.22 µm) for cell imaging to reduce background.
• Protect solutions from light during probe incubation.

These are general practices; do not substitute for item-specific instructions. Validate compatibility with SPiDER-βGal-1 using small-scale tests.

Green Alternatives

Context: Fluorogenic enzyme probes like SPiDER-βGal-1 are specialty reagents; “green chemistry” considerations primarily involve solvent choice, concentration, and waste minimization rather than replacing the probe itself.

Greener handling strategies (general):

• Prefer aqueous buffers with minimal DMSO cosolvent (≤0.1–0.5% v/v) while maintaining probe solubility and performance.
• Use microplate and microvolume formats to reduce reagent consumption and waste.
• Employ low-light, amber labware to extend probe lifetime and reduce repeated syntheses/purchases.
• Consolidate washes and use closed containers to limit VOC exposure and evaporation.

Solvent comparison for stock solutions (general):

• DMSO (common choice): miscible with water; relatively low toxicity; facilitates live-cell compatibility at low percentages.
• Alternatives (when compatible): glycerol or PEG-400/water mixtures can occasionally serve as cosolvents for more sustainable handling but may alter cell permeability or enzyme activity; validate empirically.

Waste and EHS:

• Collect spent probe solutions and rinses as hazardous organic waste; avoid drain disposal. Use enzymatic quench (add β-gal inhibitor or denature protein) before disposal to minimize bioactive components in waste.

Note: There are no direct “green alternative” probes specified for SPiDER-βGal-1 in this listing. Optimize protocols (lower probe loadings, shorter incubations, efficient imaging) to reduce environmental footprint while maintaining data quality.

Pharmaceutical Uses

This product is a research-use-only fluorescent probe for detecting β-galactosidase activity. No pharmacopeial status, excipient role, or therapeutic application is provided in the Product Data.

General considerations (non-clinical, for R&D workflows only):

• Potential roles in preclinical research include reporter-based assays for gene expression (lacZ), high-content screening of modulators of lysosomal function, or senescence markers in cell culture models. These are laboratory research applications and not pharmaceutical uses.
• Formulation insights for lab use: If formulating into assay kits or screening panels, consider aliquoting under low light into amber vials, with inert headspace and desiccant, and provide validated solvent/compatibility data to end users. Ensure all materials are labeled “For research use only.”

Compliance note:

• Do not use in humans or animals for therapeutic, diagnostic, or clinical purposes. No GMP, USP/NF, EP, or JP status is implied or stated for SKU S1455228.

For any intended regulated use or kit incorporation, obtain batch-specific CoA/Spec Sheet and SDS, and perform in-house qualification of purity, identity, performance characteristics, and stability under proposed storage/transport conditions.

Physical Properties

Item-specific physical properties:

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Purity/Grade: Not specified for this item; refer to CoA/Spec Sheet.
• Melting point, boiling point, density, refractive index, pKa, logP: Not specified for this item; refer to CoA/Spec Sheet.
• Spectral properties (λex/λem, ε, ΦF): Not specified for this item; refer to CoA/Spec Sheet.

General expectations for SPiDER-type β-gal probes (literature context only):

• Physical state: often supplied as a solid or film; highly photosensitive; commonly prepared as concentrated stock solutions in anhydrous DMSO.
• Solubility profile: good solubility in polar aprotic solvents (DMSO, DMF); working solutions typically prepared by diluting DMSO stocks into aqueous buffers (e.g., PBS, HEPES). Aqueous solubility of the intact probe can vary with formulation and counterion.
• Photophysical behavior: non-fluorescent or weakly fluorescent in the masked (β-galactoside) form; upon β-galactosidase cleavage and subsequent intramolecular trapping, a strongly fluorescent cationic dye is generated. Exact spectral maxima depend on the specific SPiDER-βGal variant.

Important: Do not treat these generalities as specifications for S1455228. For experiments requiring precise physical or spectral values, please refer to the item-specific CoA/Spec Sheet and verify with in-house QC measurements where necessary.

Quality and Grades

Item-specific quality information:

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

Interpretation and expectations (general guidance for fluorescent probes):

• Research-grade fluorescent probes are typically qualified for analytical and imaging workflows rather than for GMP uses. Key QC attributes often include identity (NMR/HRMS), purity (HPLC/UPLC), residual solvents, and verification of functional response (e.g., fluorescence turn-on upon enzymatic activation). These details are item-specific and should be confirmed in the CoA.
• Stabilizers/additives: Some photoactive probes are formulated with stabilizers or as salts to enhance stability. If any stabilizers are present, they will be declared on the Spec Sheet. Absent such declaration, do not assume their presence.
• Photostability and signal performance are sensitive to storage and handling. Protect from light and avoid repeated freeze–thaw cycles to maintain quality.

What is specific to this listing:

• Storage conditions are specified: store at −20°C, protected from light; shipped cold. Grade and exact purity are not listed for SKU S1455228. For method development (e.g., quantitative imaging or enzymology), consult the CoA/Spec Sheet for batch-specific specifications before use.

Reaction and Applications

Applicability to classical synthetic reactions: This product is a β-galactosidase-activatable fluorescent probe for research workflows, not a general-purpose synthetic reagent. It is typically used in biochemical assays and imaging rather than in stoichiometric or catalytic organic reactions.

Research applications (general, literature-based context):

• Enzyme activity detection: Fluorogenic readout of β-galactosidase activity. Hydrolysis of the β-galactoside unmasking group triggers a rapid intramolecular process that generates a bright, cationic dye.
• Cellular imaging: After activation, SPiDER-type dyes can become locally trapped, enhancing signal retention and spatial contrast in cells expressing β-galactosidase (e.g., lacZ reporter systems) or exhibiting β-gal activity associated with specific biological states.
• Plate-based assays and flow cytometry: Turn-on fluorescence facilitates high-throughput screening or single-cell analysis when paired with appropriate excitation/emission settings (exact maxima are probe-variant dependent).
• Kinetic measurements: Time-resolved fluorescence increase can be used to estimate apparent enzymatic rates under defined conditions. Calibration with known enzyme concentrations is recommended.

Practical tips (general):

• Prepare fresh DMSO stocks; minimize light exposure during preparation and incubation.
• Include no-enzyme and inhibitor controls to confirm β-gal–dependent activation.
• Validate pH and temperature for the enzyme source. Enzyme activity profiles vary widely among bacterial, mammalian lysosomal, and senescence-associated β-galactosidases.

Note: The above expands on typical manufacturer-stated uses for SPiDER-class β-gal probes. Item-specific validated applications for SPiDER-βGal-1 are not provided here; consult the CoA/Spec Sheet.

Reaction Conditions

Item-specific tested conditions: Not specified for this item; refer to CoA/Spec Sheet for any validated protocols.

General conditions for β-galactosidase activation assays (literature guidance; adjust empirically):

• Stock solution: Prepare fresh in anhydrous DMSO under low light; typical lab stocks are 1–10 mM. Filter if particulates are present. Store aliquots at −20°C, protected from light.
• Working concentration: Often in the submicromolar to low micromolar range for live-cell imaging; exact values are probe- and system-dependent.
• Temperature and time: Incubate at 20–37°C for 10–60 min, monitoring fluorescence increase over time. Optimize based on enzyme abundance and compartmental access.
• Buffer and pH: Match buffer to the enzyme source (neutral for lacZ, acidic for lysosomal, mildly acidic for SA-β-gal). Maintain physiological osmolarity for live-cell assays.
• Controls: Include no-enzyme controls, heat-inactivated enzyme controls, and known β-gal inhibitors to verify specificity. For plate assays, include a standard curve with purified β-gal when quantitative activity is required.
• Readout: Use instrument settings compatible with the expected emission channel for your SPiDER-βGal variant; verify with the CoA/Spec Sheet or by measuring excitation/emission spectra of an activated sample.

Caveats:

• Avoid high DMSO (>0.5% v/v) in live cells to minimize toxicity.
• Protect all solutions from ambient light to prevent background increase from photolysis.

These conditions are general best practices and not validated specifications for SPiDER-βGal-1.

Safety and Handling

GHS information for this item:

• Signal Word: Not specified for this item; refer to SDS.
• Hazard Statements (H-phrases): Not specified for this item; refer to SDS.
• Pictograms / GHS Classification: Not specified for this item; refer to SDS.

General safety guidance (professional laboratory practice):

• PPE: Use appropriate laboratory PPE including lab coat, safety glasses, and nitrile gloves. Handle within a fume hood when preparing concentrated stocks to minimize inhalation/dermal exposure.
• Light sensitivity: Protect from light during handling, aliquoting, and storage to prevent photodegradation. Use amber vials or wrap containers with aluminum foil.
• Incompatibilities: Avoid strong oxidizers and strong acids/bases that could hydrolyze or degrade glycosidic linkages or dye scaffolds. Minimize exposure to moisture when handling solid or concentrated DMSO stocks.
• First-aid overview: In case of skin contact, wash with soap and water; for eye exposure, rinse cautiously with water for several minutes and seek medical attention; if inhaled, move to fresh air; if ingested, rinse mouth and consult medical services. Always consult the SDS for authoritative instructions.
• Waste: Collect solutions and rinses as hazardous organic waste per institutional and local regulations. Avoid release to the environment.

Storage and shipping (from Product Data):

• Storage: Store at −20°C, protected from light.
• Shipping: Ice chest with ice pads.

Research use: For research use only. Not for human or animal therapeutic or diagnostic use.

Solvent Selection

Item-specific solvent data: Not specified for this item; refer to CoA/Spec Sheet.

General solvent guidance for SPiDER-type β-gal probes (literature/context):

• Stock preparation: Anhydrous DMSO is commonly used to prepare concentrated stocks (e.g., 1–10 mM) due to good solubility of masked fluorogenic probes in polar aprotic media.
• Working solutions: Dilute DMSO stocks into aqueous buffers (PBS, HEPES, carbonate, or acetate buffers) appropriate to the intended β-galactosidase source (bacterial lacZ vs. mammalian lysosomal vs. senescence-associated activity). Maintain low final DMSO (≤0.1–0.5% v/v) to minimize biological perturbation.
• Miscibility profile (general): DMSO is miscible with water; DMF likewise, but DMSO is typically preferred for biocompatibility. Avoid alcohol-rich media that may alter membrane permeability or enzyme activity in cell-based assays.
• pH considerations: β-Galactosidase activity is pH-dependent; buffer selection (acetate pH ~4–5 for lysosomal, phosphate/citrate pH ~6–6.5 for SA-β-gal, phosphate/HEPES pH ~7–7.4 for lacZ) should be matched to the biological context. Verify compatibility with your specific enzyme source.

Comparison (general):

• DMSO: high solubilizing power, low volatility, cell-compatible at ≤0.5% v/v.
• DMF: similar solvency; less commonly used in live-cell contexts.
• Aqueous-only: limited probe solubility and stability; typically unsuitable without a DMSO cosolvent.

Note: These are general recommendations for class-related probes; confirm solvent compatibility for SPiDER-βGal-1 with the item’s CoA/Spec Sheet and pilot tests.

Storage and Reconstitution

Storage (from Product Data):

• Store at −20°C, protected from light.
• Shipped on ice (ice chest + ice pads).

Item-specific reconstitution details: Not specified for this item; refer to CoA/Spec Sheet.

General reconstitution guidance for SPiDER-type β-gal probes (context; not product specifications):

• Solvent: Prepare concentrated stocks in anhydrous DMSO under subdued light. Vortex gently to dissolve; brief sonication may assist if needed.
• Aliquoting: Dispense single-use aliquots into amber microvials to avoid repeated freeze–thaw cycles. Flush headspace with inert gas (e.g., argon) if long-term storage is anticipated.
• Working solutions: Dilute the DMSO stock into pre-warmed, pH-appropriate assay buffer immediately before use. Keep final DMSO low (≤0.1–0.5% v/v) for live-cell studies.
• Light protection: Use amber tubes and foil wrapping during handling and incubations.

Stability and handling notes:

• Minimize freeze–thaw cycles; thaw on ice, use promptly, and refreeze unopened aliquots as soon as possible.
• Discard solutions showing precipitation, unexpected color change, or elevated background fluorescence in no-enzyme controls.

These general practices support consistent performance but are not a substitute for the item’s CoA/Spec Sheet, which should be consulted for any batch-specific instructions.

Structure and Identity

Item-specific identifiers:

• SKU: S1455228
• Product Name: SPiDER-βGal-1
• CAS: 1824699-57-1
• 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.
• CID: Not specified for this item; refer to CoA/Spec Sheet.

General structural features (literature, for context only):

• SPiDER-type β-galactosidase probes typically comprise a β-galactopyranoside moiety linked through a self-immolative/phenolic trigger to a xanthene- or coumarin-like fluorophore precursor. Upon enzymatic hydrolysis of the β-galactoside, an intramolecular reaction produces a cationic fluorophore that can form a localized, trapped species in cells.
• Functional groups commonly encountered include: glycosidic β-linkage (galactose), phenolic carbamate or ether triggers, and an electron-rich aromatic core capable of forming a highly fluorescent delocalized cation after activation.
• 2D description (literature): a galactose ring (pyranose) attached via an O-β-glycosidic bond to an aromatic linker on a latent fluorophore scaffold; the post-activation dye typically contains fused aromatic rings with dialkylamino substituents.

Notes:

• The above “general features” summarize common chemotypes reported for SPiDER-βGal probes and are not a substitute for the exact structure of SPiDER-βGal-1. For definitive identity information, consult the CoA/Spec Sheet for this specific item.

Synthetic Utility

Relevance to synthetic chemistry: SPiDER-βGal-1 is supplied as a finished fluorogenic probe and is not typically used as a building block or reagent in general organic synthesis.

General synthetic considerations (context only):

• β-Gal probes of the SPiDER class arise from convergent synthesis of a masked fluorophore (often a xanthene- or related chromophore precursor) and a β-D-galactopyranoside leaving group installed via glycosylation. The linker is engineered to undergo self-immolative cleavage and intramolecular trapping post-enzymatic activation.
• Functional groups that govern reactivity include: glycosidic β-linkage (enzymatically cleavable), phenolic/carbamate triggers (self-immolative), and electron-donating dialkylamino substituents (enhancing fluorescence upon cation formation).

If adaptation is desired (advanced users):

• Derivatization at the linker or dye periphery can tune spectral properties or subcellular localization (e.g., cationic motifs to enhance mitochondrial retention). Such modifications require de novo synthesis and full revalidation of enzyme response.

Note: The above addresses general retrosynthetic logic for the probe class. It does not imply that SPiDER-βGal-1 is intended or suitable for synthetic transformations. For this catalog item, treat it as an assay reagent, not a synthetic intermediate.

Target Specificity

Item-specific target information is not provided in the Product Data for SKU S1455228.

Per catalog constraints for this section, only product-data-derived attributes can be listed. For SPiDER-βGal-1, no validated target species, clone, isoform selectivity, or cross-reactivity data are supplied here. Refer to the CoA/Spec Sheet and SDS for any available characterization.

General note: β-Galactosidase activity can originate from multiple sources (bacterial lacZ, mammalian lysosomal, or senescence-associated activity). If target specificity (e.g., organism, isoform, cellular compartment) is critical to your study, conduct appropriate controls and consult peer-reviewed literature for assays using the exact SPiDER-βGal-1 variant.