NGF Mouse Pre-designed siRNA Set A

Cat. No.: N1459946

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Visão geral

NGF Mouse Pre-designed siRNA Set A contains three designed siRNAs for NGF gene (Mouse), as well as a negative control, a positive control, and a FAM-labeled negative control. Components NGF siRNA-1: 5 nmol (HPLC) NGF siRNA-2: 5 nmol (HPLC) NGF siRNA-3: 5 nmol (HPLC) siRNA Negative Control: 5 nmol (HPLC) FAM-labeled siRNA Negative Control: 5 nmol (HPLC) GAPDH siRNA Positive Control:5 nmol (HPLC)

Especificações

Condições de armazenamento de armazenamento

Store at -20°C

Enviado em

Ice chest + Ice pads

Este produto requer transporte de cadeia fria. Serviços terrestres e outros serviços econômicos não estão disponíveis.

Product Line

Produtos biológicos

Application Protocols

Item-specific validated applications, recommended dilutions, and step-by-step protocols: Not specified for this item; refer to CoA/Spec Sheet or the manufacturer’s protocol guide provided with the product.

Notes:

• As a general resource, follow a transfection reagent protocol optimized for siRNA and your cell type. Verify NGF knockdown by RT‑qPCR and protein assays, and include appropriate controls.
• Any protocol details such as plating density, complexation volumes, or incubation times must be established empirically for your system or obtained from the supplied instructions if available.

Biological Roles

Target biology overview (general literature for NGF in mouse):

• NGF (nerve growth factor) is a prototypical neurotrophin critical for survival, differentiation, and maintenance of sympathetic and certain sensory neurons. It signals primarily through the high-affinity receptor TrkA (NTRK1) and the low-affinity p75NTR (NGFR).
• Functions: Promotes neurite outgrowth, supports cholinergic neuron function, and modulates nociception/pain sensitization. NGF also exerts roles in immune cell function and tissue repair, with context-dependent effects in inflammation.
• Signaling: TrkA activation initiates downstream MAPK/ERK, PI3K/AKT, and PLCγ pathways; p75NTR can modulate survival/apoptosis signaling depending on ligand/receptor context.
• Expression/regulation: NGF is produced by target tissues and various non-neuronal cells; transcription is regulated by activity, cytokines, and stress. Multiple transcript variants and mature peptide processing are described in the literature.

Reagent relevance:

• An siRNA set against mouse NGF enables transient suppression of NGF mRNA to interrogate pathway dependence, gene–phenotype relationships, and compensatory mechanisms in cellular models. Researchers can correlate NGF knockdown with changes in neuronal morphology, trophic signaling readouts, or cytokine responses, as appropriate for their system. All study designs should include proper controls and verification at mRNA and protein levels.

Note: The above is general background; no performance characteristics are claimed for this specific product beyond research use.

Buffer Applications

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

General buffer practices for siRNA handling (literature):

• Stock solutions: Prepare in RNase‑free water or TE‑Low (e.g., 10 mM Tris‑HCl, pH 7.5–8.0, with 0.1–1 mM EDTA). EDTA chelates trace divalent cations that catalyze RNA cleavage, enhancing stability during storage.
• Working dilutions: Often made in cell culture medium immediately before transfection, following the transfection reagent’s protocol. Avoid high ionic strength stocks that may promote aggregation.
• RNase control: Use RNase‑free consumables; decontaminate benches and equipment with RNase‑inactivating solutions. Avoid repeated freeze–thaw cycles by preparing single‑use aliquots.
• Sterility: If sterile solutions are required, prepare under aseptic conditions and/or pass through a 0.22 µm RNase‑free, low‑binding filter; do not heat‑sterilize RNA.

These are general guidelines for oligonucleotide buffers and not item-specific specifications.

Green Alternatives

Context: As an aqueous, nonvolatile oligonucleotide reagent, siRNA already avoids many environmental burdens associated with organic solvents and volatile reagents.

Greener practice considerations (general):

• Solvent profile: Use RNase‑free water or TE‑Low for stock solutions; minimize or eliminate organic cosolvents. Prepare concentrated stocks to reduce plastic waste from aliquoting.
• Delivery systems: Prefer transfection reagents with documented lower toxicity and improved biodegradability; verify manufacturer environmental data.
• Waste minimization: Plan experiments to scale appropriately, use multiwell formats, and pool replicates to reduce reagent consumption.
• Packaging and cold-chain: Consolidate orders to reduce shipments; promptly transfer to −20 °C to minimize dry ice/ice pack waste.

Comparison to small-molecule inhibitors (conceptual):

• Specificity: Sequence-directed knockdown can reduce the need for panels of chemical inhibitors with varied off-target profiles.
• Persistence: siRNA effects are transient, potentially reducing long-term environmental persistence relative to some halogenated organics.

All recommendations above are general sustainability practices; they do not alter the storage, safety, or performance requirements of this specific item.

Pharmaceutical Uses

Regulatory/compendial status for this item: Not specified; this product is labeled for research use only.

Context (general, non-clinical):

• siRNA reagents like this are commonly employed in preclinical research to validate targets, map pathways, and inform medicinal chemistry or biologic development by transiently reducing gene expression.
• Formulation learnings: Research-formulated siRNA complexes (e.g., lipid nanoparticles, polymer conjugates) in discovery settings can inform later translational strategies; however, this product is not manufactured or released under GMP, and is not intended for diagnostic, therapeutic, or in vivo clinical use.
• Excipient compatibility: Any excipient interactions or stability profiles relevant to pharmaceutical development must be determined empirically for the specific sequence and formulation and are outside the scope of this research reagent.

No therapeutic or clinical claims are made. For any use beyond laboratory research, consult appropriate regulatory frameworks and procure GMP-grade materials.

Physical Properties

Item-specific physical specs (appearance, concentration, solvent, extinction coefficients): Not specified for this item; refer to CoA/Spec Sheet.

General properties for siRNA duplexes (literature/computed context; sequence- and salt-dependent):

• Physical state: Commonly supplied lyophilized or in RNase‑free aqueous buffer; forms clear solutions upon reconstitution.
• Solubility: Readily soluble in RNase‑free water or low-ionic strength buffers (e.g., 10 mM Tris, 0.1–1 mM EDTA) to ≥100 µM (literature); actual maximum concentration depends on sequence and length.
• pH stability: Stable near neutral pH; hydrolysis accelerates at high pH (>9) due to 2'‑OH transesterification (literature).
• Temperature sensitivity: Duplex integrity maintained on ice/4 °C short term; long-term storage recommended at −20 °C or below to limit hydrolysis and depurination (literature).
• UV absorbance: Strong at 260 nm; A260 varies with sequence and concentration; OD260 to nmol conversion is sequence-dependent (literature). Not specified for this item; refer to CoA for exact OD factors if provided.
• Melting temperature (Tm): Typically 55–85 °C for 19–21 bp siRNA in ~100 mM monovalent salt; varies with GC content and modifications (literature).

Density, boiling/melting point, logP, refractive index: Not applicable to oligonucleotide aqueous solutions; not specified for this item.

Quality and Grades

Item-specific grade/purity and purification method: Not specified for this item; refer to CoA/Spec Sheet.

Context for siRNA quality (general/literature):

• Common purification levels: Desalted, cartridge/IE-HPLC purified, or PAGE purified. HPLC/PAGE typically yield higher duplex purity and narrower length distribution, which can improve knockdown consistency.
• Identity/Content verification: MALDI‑TOF/ESI‑MS for intact mass, analytical HPLC for purity, UV/OD260 for quantitation. Some vendors provide strand stoichiometry and annealing verification.
• Modifications: Chemical stabilization (e.g., 2'‑OMe, 2'‑F, phosphorothioate ends) can enhance nuclease resistance; any such features must be specified by the manufacturer.
• Endotoxin/bioburden: For cell culture use, low endotoxin levels are often targeted. Specification, if any, must come from the item CoA.
• What “Pre-designed set” implies (general): Multiple duplexes against distinct regions of the target transcript, allowing selection of the most effective sequence and controlling for off-targets.

Please consult the CoA/Specification Sheet packaged with the product for the exact purification, purity, strand length, modification pattern, and quantitation method applicable to SKU N1459946.

Reaction and Applications

Manufacturer-stated applications: Not explicitly provided for this item; siRNA sets are generally used for gene silencing in research.

Functional use (general/literature):

• RNA interference (RNAi): siRNA duplexes guide RISC to complementary NGF mRNA in mouse cells, resulting in sequence-specific mRNA cleavage and knockdown of NGF expression.
• Experimental contexts: transient knockdown in cell culture (e.g., primary neurons, fibroblasts, immune cells), organotypic cultures, and in vivo research when formulated with appropriate delivery systems (note: formulation and in vivo use require method development and institutional approvals).
• Set utility: Using multiple predesigned siRNAs permits selection of the most effective duplex and reduces the probability that observed phenotypes arise from off-target activity.

Practical notes (literature/practice):

• Titration: Optimize siRNA and transfection reagent amounts for each cell type; excessive siRNA can increase off-target effects and cytotoxicity.
• Controls: Include non-targeting siRNA, positive control siRNA, and, where possible, rescue experiments (e.g., NGF cDNA resistant to siRNA) to confirm specificity.
• Readouts: RT‑qPCR for mRNA reduction, Western blot/ELISA for NGF protein levels, and phenotypic assays relevant to NGF biology.
• Timing: Maximal knockdown typically 24–72 h post-transfection; duration depends on cell division rate and siRNA stability (literature).

Reaction Conditions

Chemical reaction conditions are not applicable. Below are general experimental conditions for siRNA-mediated knockdown (literature/practice guidance; not item-specific):

• Cell types: Optimize per cell line/primary culture. Adherent lines often transfect readily; primary neurons may require neuron-optimized reagents or electroporation.
• siRNA concentration: Common screening ranges are 1–50 nM final; many lines respond well at 5–20 nM. Start with a small matrix to balance efficacy and off-target risk.
• Transfection reagent: Choose a reagent validated for siRNA; follow vendor ratios. Complexation typically proceeds in serum-free medium for 5–20 min at room temperature.
• Medium/Serum: Many reagents allow transfection in the presence of serum after complex formation; verify compatibility to preserve cell health.
• Incubation: Replace medium after 4–24 h if cytotoxicity is observed. Assess knockdown at 24, 48, and 72 h to capture kinetics.
• Controls: Include non-targeting siRNA, a toxic siRNA control (optional), and, if available, a siRNA known to silence a housekeeping gene to verify transfection competence.
• Readouts: RT‑qPCR for NGF mRNA; Western/ELISA for NGF protein; downstream signaling assays (e.g., phospho‑TrkA, ERK) as appropriate.

For in vivo or ex vivo delivery, conditions are model- and formulation-dependent and require separate optimization and ethics approvals.

Safety and Handling

GHS/SDS status for this item:

• Signal word, hazard statements, pictograms, and classification: Not specified for this item; refer to SDS.

General safety for synthetic siRNA reagents (informational, not a substitute for the SDS):

• Expected hazards: siRNAs are nonvolatile, nonflammable aqueous/solid biomolecules and are generally considered low hazard; main risks relate to dust generation (if lyophilized), potential irritation, and contamination with RNases or microbial agents if mishandled.
• PPE: Wear lab coat, safety glasses, and appropriate gloves (nitrile). Avoid generating aerosols or dust during reconstitution. Use RNase‑free technique.
• Handling: Work in a clean area dedicated for RNA. Use RNase‑free tubes, tips, and buffers. Change gloves frequently to avoid RNase introduction. Avoid repeated freeze–thaw cycles that can degrade RNA.
• Incompatibilities: Strong bases (promote RNA transesterification), RNases (enzymatic degradation), and nucleases in general. Avoid prolonged exposure to elevated temperatures and metal-ion–rich solutions lacking chelators.
• First aid (general): If contact with eyes/skin, rinse with water. If inhaled (powder), move to fresh air. If ingested, rinse mouth and seek medical advice. Always follow the SDS for authoritative guidance.
• Spill/Disposal: Wipe small spills with RNase-decontaminating solution followed by water. Dispose according to institutional and local regulations for nonhazardous biological laboratory waste.

Solvent Selection

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

General guidance for siRNA dissolution (literature/practice):

• Primary solvent: RNase‑free water is typically used to prepare concentrated stocks (e.g., 10–100 µM). TE or TE‑Low (10 mM Tris, 0.1–1 mM EDTA, pH ~7.5–8.0) can provide chelation of trace divalent cations that catalyze RNA cleavage.
• Compatibility: siRNA duplexes are hydrophilic and not soluble in nonpolar organic solvents. Avoid alcohol-rich environments during long-term storage as they can alter hydration and duplex behavior.
• Buffer considerations: Maintain near-neutral pH. Avoid strong bases; avoid Mg2+/Mn2+ without chelators unless immediately proceeding to complexation with delivery systems.
• Filtration: Use low-protein-binding, RNase‑free filters if sterilization is required. Do not heat to sterilize.

Comparison (literature):

• RNase‑free water: maximal simplicity, compatible with most transfection mixes; lower ionic strength may increase adsorption losses.
• TE/TE‑Low: improves stability during storage by chelating metals; verify compatibility with your transfection reagent, as EDTA can impact some formulations.
• PBS and high-salt buffers: generally avoided for stock solutions to minimize duplex aggregation; can be used after dilution in certain delivery systems if validated.

Storage and Reconstitution

Item-specific storage and shipping (from Product Data):

• Storage Conditions: Store at −20 °C.
• Shipped In: Ice chest + Ice pads.

Item-specific details on form, concentration, and reconstitution volumes: Not specified for this item; refer to CoA/Spec Sheet.

General guidance (informational):

• Upon receipt: Inspect packaging temperature and integrity. Centrifuge briefly to collect material in tube bottom before opening.
• Reconstitution: Use RNase‑free water or TE‑Low to prepare concentrated stocks. Mix gently without vortexing to minimize shearing and foaming.
• Aliquoting: Prepare single‑use aliquots to avoid freeze–thaw. Label aliquots with sequence/set ID, concentration, and date.
• Storage: Keep working solutions at 4 °C for short term (≤1 week) if needed; for long term, store at −20 °C or below. Avoid frost-free freezer cycles. Protect from repeated temperature fluctuations.
• Stability: Actual shelf life and stability are sequence- and formulation-dependent; defer to CoA for any provided expiry dating or stability testing.

For authoritative instructions on reconstitution volumes, final buffer composition, and calculated nmol/OD values, consult the CoA/Specification Sheet supplied with SKU N1459946.

Structure and Identity

Item-specific identifiers (CAS, SMILES, InChIKey, exact sequence, modifications): Not specified for this item; refer to CoA/Spec Sheet.

Overview (general/literature for siRNA):

• This product is described as NGF Mouse Pre-designed siRNA Set A, indicating a collection of short interfering RNA duplexes designed to silence the mouse NGF gene (nerve growth factor) via RNA interference.
• Typical siRNA architecture (literature): 19–21 bp duplex with 2-nt 3' overhangs on each strand; sense strand matches mRNA sequence; antisense (guide) strand is complementary to the target.
• Chemical features (literature): Ribose backbone with 2'‑OH (unless chemically modified), phosphodiester (or phosphorothioate in modified designs) linkages, terminal phosphate states may vary after synthesis/purification; may be delivered lyophilized or in RNase‑free solution.
• Stereochemistry: Nucleic acids possess defined chiral centers on ribose; overall duplex adopts A‑form helical geometry in aqueous conditions (literature).
• Molecular formula and exact molecular weight: Sequence‑dependent; not defined for this multi‑duplex set. Not specified for this item; refer to CoA/Spec Sheet for strand lengths, sequences, modifications, and MW per duplex.

Target identity (from product name):

• Target gene: NGF (mouse). Ensemble/RefSeq accession numbers and target exon positions are not provided here; consult the CoA or design sheet if supplied with the set.

Synthetic Utility

This product is an siRNA reagent, not a small-molecule building block; classical organic synthetic transformations (e.g., cross-couplings, oxidations) are not applicable.

Research utility (general/literature):

• Functional genomics tool: Enables loss-of-function studies to probe NGF involvement in cellular phenotypes, signaling, and gene regulatory networks.
• Network analysis: Combining NGF knockdown with transcriptomics/proteomics can delineate downstream effectors and compensatory neurotrophin pathways.
• Assay development: Can serve as a positive control to benchmark assay responsiveness to NGF pathway suppression when developing screening campaigns.

If chemical conjugation (e.g., fluorophores, cholesterol) or stabilization chemistries are desired, such features must be specified at purchase; none are stated for this item. For custom synthetic needs, refer to custom oligonucleotide services.

Target Specificity

From product name and category:

• Target gene: NGF (nerve growth factor).
• Species: Mouse.
• Format: Pre-designed siRNA Set A.

Sequences, number of duplexes, target exon regions, and predicted off-target analysis: Not specified for this item; refer to CoA/Spec Sheet.

General considerations (informational):

• Predesigned sets typically include multiple siRNA duplexes targeting distinct regions of the NGF transcript to maximize the likelihood of effective knockdown and to enable concordance checks across sequences.
• Specificity validation should include demonstration of mRNA/protein reduction without broad cytotoxicity and lack of effect with non-targeting controls.

Perguntas frequentes

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.