Filipin complex ready made solution

Cat. No.: F755446

QUALITÀ & PUREZZA from Streptomyces filipinensis, 5mg/ml in DMSO based solution

Panoramica

Filipin Complex, the collective name given to family of isomeric polyene macrolides isolated from cultures of S. filipinensis, with Filipin III being the majority part of the complex.

Application:

Filipin complex readymade solution has been used in the staining of HeLa cells for cytoskeletal imaging.

Filipin has been used in a double staining procedure as a probe for the detection of lipoproteins in polyacrylamide gel and immobilized on nitrocellulose membranes. It is also widely used to localize and quantitate unesterified cholesterol by virtue of a specific fluorescent complex.

Specifiche

Product Name

Filipin complex ready made solution

Specifiche e purezza

from Streptomyces filipinensis, 5mg/ml in DMSO based solution

Meccanismi biochimici e fisiologici

Filipin is an antibiotic with antifungal functionality. Filipin complex has been found to alter cell membrane structure by interacting with the membrane sterols ergosterols and cholesterol. On binding to cholesterol, filipin shows fluorescence shift. As s

Tipo di molecola

Bioreagenti/Tampone

Stoccaggio e spedizione

Concentrazione

from Streptomyces filipinensis, 5mg/ml in DMSO based solution

Condizioni di conservazione di stoccaggio

Store at -20°C

Spedito in

Ice chest + Ice pads

Application Protocols

Tested applications, validated dilutions, and step‑by‑step procedures for this specific item are Not specified for this item; refer to CoA/Spec Sheet and product datasheet.

General protocols are widely available in the literature for filipin‑based cholesterol staining in cells and tissues; consult peer‑reviewed methods and adapt to your instrumentation and biosafety framework.

Biological Roles

General biological context for filipin (literature; not item‑specific):

• Binding target: Filipin binds 3β‑hydroxysterols via noncovalent complexation, inserting into lipid bilayers and associating with the sterol ring system. Cholesterol is the principal cellular target; ergosterol and other 3β‑OH sterols are also bound with varying affinity.
• Fluorescence mechanism: Complexation with sterols restricts filipin’s polyene motions and alters local polarity, enhancing blue fluorescence (emission ~480–500 nm) upon UV excitation (~340–365 nm). Signal intensity reflects local sterol accessibility and microenvironment rather than absolute concentration.
• Cellular distribution: Staining commonly highlights plasma membrane and cholesterol‑rich intracellular membranes; after fixation, filipin reports unesterified cholesterol pools. Esterified cholesterol in lipid droplets is poorly labeled without prior hydrolysis.
• Biological caveats: Filipin can perturb membranes at higher concentrations or prolonged exposure and may be cytotoxic. It photobleaches readily; imaging must be optimized to limit UV dose. Serum proteins can sequester filipin or sterols and reduce signal.
• Controls and specificity: Sterol depletion (e.g., cyclodextrin extraction) diminishes filipin labeling; addition of exogenous cholesterol can restore/enhance it. Compounds that oxidize or esterify cholesterol alter filipin staining patterns accordingly.

These roles summarize how filipin is leveraged as a research probe for membrane sterol biology. They are not clinical/diagnostic claims and do not imply performance characteristics for this specific product lot.

Buffer Applications

Filipin is not a buffering agent. However, buffer choice strongly influences staining outcomes (general literature guidance; not item‑specific):

• Staining buffers: Use serum‑free, protein‑free buffers (e.g., PBS or HBSS without phenol red) during staining to minimize sterol extraction/binding by serum proteins and to reduce background autofluorescence.
• Fixation: When staining fixed cells or tissues, avoid alcohol‑based fixatives that extract sterols. Paraformaldehyde (PFA, e.g., 3–4%) in PBS is commonly used, followed by thorough washes to remove residual fixative.
• Permeabilization: Mild, nonionic detergents (e.g., saponin) can increase access to intracellular membranes; over‑permeabilization may redistribute sterols and increase background.
• Additives: BSA (low %) may reduce nonspecific adsorption to plastics but can also sequester filipin; validate in your system.
• Light management: Perform incubations and washes under low light; use amber tubes when possible.

Note: The exact buffer composition, incubation times, and temperatures suitable for this item are Not specified for this item; refer to CoA/Spec Sheet and your lab’s validated protocols.

Green Alternatives

While filipin is a specialized sterol probe (with few functional substitutes providing identical readouts), users can consider alternatives that reduce solvent hazards, phototoxicity, or waste generation. The choice depends on assay goals.

Comparison (literature/context; not item‑specific):

• Filipin complex

  • Strengths: direct, label‑free binding to 3β‑hydroxysterols; bright blue emission; fast staining.
  • Tradeoffs: UV excitation (phototoxic, autofluorescence), photobleaching, organic solvent carriers (DMSO/EtOH), limited quantitative linearity.

• Protein‑based probes (e.g., perfringolysin O D4 domain, D4H sensors)

  • Strengths: visible‑range fluorescent tags; improved specificity for accessible cholesterol; live‑cell compatibility with genetic expression.
  • Tradeoffs: cloning/expression required; potential perturbation of membranes; larger biomolecules, more waste from biological prep.

• Synthetic fluorescent sterols (e.g., BODIPY‑cholesterol, dehydroergosterol)

  • Strengths: visible excitation (BODIPY), better photostability (some analogs), can enable quantitative uptake studies.
  • Tradeoffs: probe behaves as sterol analog (not native cholesterol), different partitioning/kinetics; may require specialized optics.

Greener handling tips for filipin (general):

• Use the lowest effective working concentration and minimize organic co‑solvent in final media.
• Employ amber vials and LED illumination to reduce photodegradation and repeat trials.
• Batch experiments to reduce thaw cycles and waste.

Select the probe/approach that balances data needs with solvent load, illumination requirements, and biosafety in your lab.

Pharmaceutical Uses

This product is supplied for research use only.

Context (general information; not item‑specific and not a clinical claim):

• Filipin is a polyene macrolide complex historically characterized as an antifungal natural product; however, in the research laboratory it is primarily used as a fluorescent analytical probe for sterols rather than as a formulated excipient or process reagent.
• It does not have a recognized role as a pharmaceutical excipient, and UV excitation requirements and photolability limit its use outside analytical contexts.

Manufacturing/formulation relevance: none typical. For any use beyond laboratory research, consult appropriate regulatory guidance. No pharmacopeial monograph applies to this ready‑made solution to our knowledge.

Item‑specific regulatory/grade information: Not specified for this item; refer to CoA/Spec Sheet.

Physical Properties

• Item-specific solution properties (solvent, concentration, appearance): Not specified for this item; refer to CoA/Spec Sheet.
• Density, refractive index, pH (if aqueous), and UV cutoff for the supplied solution: Not specified for this item; refer to CoA/Spec Sheet.

Literature/typical properties of the Filipin fluorophore (not specifications for this item):

• Chromophore: polyene macrolide; strongly absorbs in the near‑UV due to conjugation.
• Excitation/emission (literature, in cells or organic media):
  • Excitation typically ~340–365 nm
  • Emission typically ~480–500 nm (blue fluorescence)
  • Fluorescence is enhanced upon binding to 3β‑hydroxysterols (e.g., cholesterol) in membranes; signal is sensitive to environment and photobleaches relatively rapidly.
• Solubility (literature): sparingly soluble in water; soluble in polar organics (e.g., DMSO, DMF, ethanol). Working solutions are commonly prepared in DMSO or ethanol and diluted into aqueous buffers immediately before use to minimize precipitation.
• Stability (literature): light‑sensitive; prone to photobleaching and slow oxidative degradation. Improved stability when stored cold and protected from light; avoid repeated freeze–thaw.

Practical notes (general guidance):

• Use low‑autofluorescence plastics/glass for UV/blue detection.
• Maintain minimal exposure to ambient light during handling and imaging to preserve fluorescence.

Quality and Grades

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

Context for this ready‑made solution (general guidance):

• Ready‑to‑use convenience: Pre‑dissolved filipin reduces variability from user dissolution and limits light/air exposure during preparation, improving lot‑to‑lot reproducibility in imaging assays.
• Concentration control: The working concentration and solvent identity drive staining intensity and cell compatibility. These are item‑ and lot‑specific parameters that must be verified on the CoA.
• Fluorescence performance: For microscopy or flow applications, low background and consistent emission depend on controlled impurity profiles in both filipin and solvent. HPLC/fluorescence assessment is commonly used by advanced labs; consult the CoA for any provided chromatographic purity or assay details.
• Stabilizers: If present, stabilizers (e.g., antioxidants) can slow photodegradation but may influence biological assays. Presence/identity of any stabilizer is Not specified for this item; refer to CoA/Spec Sheet.

Documentation to consult before use:

• Product CoA for solvent, concentration, lot‑specific specs, and any stabilizer.
• SDS for hazard classifications and safe handling.
• Internal method notes for imaging/flow settings to align with the supplied concentration.

Reaction and Applications

This product is not typically used as a synthetic reagent or catalyst. It is primarily a biochemical probe for sterols. Therefore, classical organic reaction applications (e.g., coupling, reductions) are not applicable here.

Relevant laboratory applications (general literature guidance):

• Fluorescent staining of unesterified cholesterol in fixed or live cells for epifluorescence or confocal microscopy (excitation ~340–365 nm; emission ~480–500 nm).
• Assessing sterol distribution in membranes, endolysosomal compartments, and model membranes (liposomes, GUVs) via filipin–sterol complex fluorescence.
• Qualitative cholesterol mapping in tissue cryosections after aldehyde fixation (ethanol‑free fixation preferred to avoid sterol extraction).
• Controls and validation: sterol depletion (e.g., methyl‑β‑cyclodextrin treatment) decreases filipin signal; sterol enrichment increases signal (conceptual controls; implement per your institutional approvals).

Note: The above are general uses from the literature and are not item‑specific performance claims. See the Application Protocols tab and the product CoA for any product‑validated uses (if provided).

Reaction Conditions

Not applicable for this product. Filipin complex ready‑made solution is a biochemical probe used for fluorescent staining rather than for performing chemical reactions.

If using in biological assays, consult your validated protocols for incubation times, temperatures (often ambient for fixed cells; 4–37°C ranges may be used depending on application), and illumination settings. Item‑specific recommended conditions are Not specified for this item; refer to CoA/Spec Sheet.

Safety and Handling

Item‑specific hazard classification (GHS), pictograms, and H‑statements: Not specified for this item; refer to SDS.

General safety guidance for filipin solutions (literature/practice; not item-specific):

• Hazards: Polyene macrolides can be irritant and harmful if swallowed, inhaled, or in contact with skin and eyes. Solutions may contain organic solvents (e.g., DMSO or ethanol), which add their own hazards (flammability/irritation). Filipin is photosensitive and may degrade to unknown products upon extended light exposure.
• PPE: lab coat, nitrile gloves, and chemical splash goggles. Handle in a well‑ventilated area or fume hood to minimize inhalation of vapors/ aerosols from the solvent.
• Handling:
  • Minimize light exposure (work under dim light or use amber containers).
  • Avoid repeated freeze–thaw; aliquot upon first thaw if the concentration permits (see CoA for concentration).
  • Prevent contamination; use clean, dry, chemically compatible tools.
• Incompatibilities (general): strong oxidizers; strong acids/bases that can degrade polyenes; light and heat accelerate decomposition. Avoid contact with reactive plastics if supplied in aggressive solvents.
• First aid (overview; defer to SDS):
  • Skin/eye contact: rinse with water for at least 15 minutes; remove contaminated clothing; seek medical attention if irritation persists.
  • Inhalation: move to fresh air; seek medical attention if symptoms occur.
  • Ingestion: rinse mouth; do not induce vomiting; seek medical advice.
• Waste: Collect as halogen‑free organic waste or mixed solvent waste per your institution’s protocols; shield from light in waste container; label contents clearly.

Always consult the product‑specific SDS for authoritative hazard, response, and disposal requirements.

Solvent Selection

Because this product is provided as a ready‑made solution, solvent choice for initial dissolution is predetermined. Item‑specific solvent identity and concentration are Not specified for this item; refer to CoA/Spec Sheet.

General solvent considerations for filipin use (literature/practice):

• Polarity/miscibility: Filipin is poorly water‑soluble but soluble in polar organics (DMSO, DMF, ethanol). Working dilutions are typically made by adding the stock slowly to aqueous buffers (e.g., PBS) with gentle mixing to avoid precipitation.
• Dielectric and partitioning: DMSO (ε ~47) affords strong solubilization and rapid membrane delivery but may affect cells at ≥0.5–1% v/v. Ethanol (ε ~24) is common but can be cytotoxic >1–2% v/v. Keep final organic content as low as compatible with staining.
• Buffer compatibility: Use serum‑free buffers during staining to avoid cholesterol sequestration by albumin/lipoproteins; add back serum after staining if required.

Comparison (literature guidance):

• DMSO stocks: highest solubility; minimal precipitation on dilution if added last and mixed well; watch for DMSO‑related biological effects.
• Ethanol stocks: adequate for many assays; faster evaporation; increased flammability; can extract membrane lipids at higher %.

Recommendation: Verify the supplied solvent on the CoA and adjust dilution protocols to maintain final organic solvent below your experimental tolerance.

Storage and Reconstitution

• Storage (item‑specific from Product Data): Store at −20°C.
• Shipping (item‑specific from Product Data): Shipped in an ice chest with ice pads.
• Light sensitivity: Filipin is light‑sensitive; keep the container protected from light (amber vial or foil wrap) during storage and handling.
• Freeze–thaw: Minimize cycles. If the stock concentration allows, prepare aliquots under low light on first thaw to avoid repeated freeze–thaw stress.
• Reconstitution: Not applicable for a ready‑made solution unless concentrated or dried; any dilution solvent/composition for working solutions is Not specified for this item; refer to CoA/Spec Sheet.
• Handling on receipt: Store promptly at −20°C upon arrival. Thaw on ice or at 4°C if needed, mix gently to homogenize, and keep cold and protected from light during setup.
• Stability after opening (general guidance): Filipin solutions can degrade with light, oxygen, and heat. Plan experiments to limit bench‑top exposure. For long experiments, keep stocks on ice and shielded from light.

For definitive shelf life, concentration, solvent, and stabilizer information, consult the lot‑specific CoA and SDS.

Structure and Identity

This product is a ready‑made laboratory solution of the Filipin complex, a polyene macrolide mixture classically used as a fluorescent probe for 3β‑hydroxysterols (e.g., cholesterol) in biological membranes.

• Item-specific identifiers:

  • CAS: Not specified for this item; refer to CoA/Spec Sheet.
  • Molecular formula: Not specified for this item; refer to CoA/Spec Sheet (note: Filipin “complex” is a mixture and does not have a single empirical formula).
  • Molecular weight: Not specified for this item; refer to CoA/Spec Sheet (mixture, component MWs vary in literature).
  • InChIKey / SMILES: Not specified for this item; refer to CoA/Spec Sheet (mixture, multiple congeners).

• Literature description (for context; not item‑specific):

  • The Filipin complex contains several closely related polyene macrolides (e.g., filipin I–IV; often dominated by filipin III). Each congener comprises a large 28–38‑membered macrolactone with multiple conjugated double bonds (polyene), numerous hydroxyl substituents, and a lactone carbonyl.
  • Structural features: macrocyclic lactone ring; extended polyene chromophore; multiple secondary alcohols; amphipathic topology enabling sterol complexation.

• 2D structure (described verbally): a large macrocyclic ring (lactone) bearing an extended, alternating double‑bond segment (polyene) along one arc, with several hydroxyl‑bearing stereogenic centers on the opposite arc; no halogens or heterocycles beyond the ester (lactone) functionality.

Note: Because this is a complex mixture provided as a solution, single-structure identifiers and exact composition are inherently mixture‑dependent and must be confirmed on the specific lot CoA.

Synthetic Utility

Filipin complex is not typically employed as a building block, reagent, or catalyst in synthetic organic chemistry. It is a labile, high‑molecular‑weight polyene macrolide used for biochemical staining, not for chemical transformations.

• Applicability to synthesis: Not applicable. The compound’s multiple conjugated double bonds and dense functionality make it unsuitable as a general synthetic reagent.

For research needs, focus on the Biological Roles, Buffer Applications, and Storage tabs rather than synthetic methodology.

Target Specificity

This section typically applies to antibodies or affinity reagents with defined antigen/epitope data. No such item‑specific target data are provided for this product.

• Item‑specific target/epitope information: Not specified for this item; refer to CoA/Spec Sheet.

Note: In general literature, filipin binds 3β‑hydroxysterols in membranes, but this is contextual background rather than a product‑validated specificity claim.

Domande frequenti

How should this product be stored?

Store at ?20 °C. Freezer storage is required to maintain the specified shelf life.

How is this product shipped?

This product ships in an insulated container with ice pads. Unpack on arrival and transfer it to the storage condition stated above.

What documentation is provided?

Available product documentation, including Certificates of Analysis (COA), Safety Data Sheets (SDS), and specification sheets, is shown in the product document area. Document availability and access follow the current site policy.