Grm4 Mouse Pre-designed siRNA Set A

Cat. No.: G1469296

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Übersicht

Grm4 Mouse Pre-designed siRNA Set A contains three designed siRNAs for Grm4 gene (Mouse), as well as a negative control, a positive control, and a FAM-labeled negative control. Components Grm4 siRNA-1: 5 nmol (HPLC) Grm4 siRNA-2: 5 nmol (HPLC) Grm4 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)

Specifications

Storage

Store at -20°C

Verschickt in

Ice chest + Ice pads

Dieses Produkt erfordert Kühlkettenversand. Grundversand und andere Economy-Optionen sind nicht verfügbar.

Product Line

biological_products

Application Protocols

Item-specific tested applications, recommended protocols, and dilutions are not provided in the Product Data for this siRNA set. Please refer to the product datasheet/CoA for authoritative instructions.

General note: While typical siRNA workflows include reconstitution, transfection optimization, and knockdown verification, such protocols are considered general literature guidance and not product-specific validation. For reproducible results with the Grm4 Mouse Pre-designed siRNA Set A, follow the vendor’s sequence-specific and reagent-specific recommendations when available.

Biological Roles

• Item-specific: The product targets mouse Grm4 by RNA interference; specific sequences and validation data are not provided here.

• Target biology (literature, Mus musculus):

  • Grm4 encodes metabotropic glutamate receptor 4 (mGluR4), a class C G protein–coupled receptor (GPCR) primarily coupled to Gi/o proteins. Activation typically inhibits adenylyl cyclase, reducing intracellular cAMP.
  • mGluR4 is predominantly expressed in the cerebellum and basal ganglia, localized to presynaptic terminals where it modulates neurotransmitter release and synaptic plasticity.
  • Endogenous ligands are glutamate and related excitatory amino acids; the receptor participates in fine-tuning excitatory neurotransmission and network excitability.
  • Signaling crosstalk: mGluR4 can influence ion channel activity and interact with scaffold proteins; downstream effects may span MAPK pathways and synaptic vesicle cycling.

• Application implications (general):

  • siRNA-mediated reduction of Grm4 enables dissection of presynaptic GPCR function, short-term plasticity, and circuit-level modulation in neuronal models.
  • Functional assays may include: cAMP accumulation, paired-pulse ratios in electrophysiology, synaptic vesicle release assays, and ligand-response profiling with group III mGluR agonists/antagonists.

Note: All biological role details above are general literature context and not product-specific performance claims.

Buffer Applications

This product is a nucleic acid tool rather than a chemical buffer. The relevant consideration is the choice of buffers for reconstituting and handling siRNA.

• Item-specific: No buffer composition is provided for this item; refer to CoA/Spec Sheet.

• General guidance (literature/practice):

  • Reconstitution: RNase-free water is suitable. For improved stability, 10 mM Tris-HCl, pH 7.4–8.0, with 0.1–1 mM EDTA (TE) is commonly used for long-term stocks.
  • Working solutions: Prepare in RNase-free water or serum-free media immediately before complexing with transfection reagents. Avoid Mg2+/Ca2+ in storage buffers; they can accelerate RNA hydrolysis.
  • Ionic strength: Low salt favors duplex stability during storage; physiological buffers are fine post-complex formation for cell exposure.
  • Annealing (if single strands supplied): Heat to 90–95 °C for 1–2 min in annealing buffer (e.g., 30 mM HEPES-KOH pH 7.4, 100 mM potassium acetate, 2 mM magnesium acetate), then cool slowly to room temperature; for this catalog item, duplex status is not specified.
  • Sterility: Filter buffers (0.22 μm) prior to contact with RNA; do not filter RNA duplexes to avoid adsorption losses.

All buffer recipes provided are general guidance; use the product documentation for item-specific instructions.

Green Alternatives

While siRNA use has a modest environmental footprint, delivery and processing steps can involve hazardous reagents. Consider the following greener practices (general/literature guidance):

• Transfection alternatives:

  • Prefer biodegradable or less toxic cationic polymers/lipids over older quaternary ammonium reagents known for cytotoxicity and persistence.
  • Use electroporation or physical delivery (microfluidic squeezing) to eliminate chemical transfection waste, where compatible with cell type.

• Buffering and plasticware:

  • Prepare RNase-free buffers in-house to reduce packaging. Choose phosphate-free buffers where eutrophication is a concern and not required by the protocol.
  • Use low-retention, recyclable plastics and minimize single-use consumables via careful planning while maintaining RNase-free technique.

• Waste minimization:

  • Scale reactions to the minimal volume that provides robust readouts. Consolidate controls across experiments.
  • Employ non-ethidium bromide nucleic acid stains (e.g., SYBR Safe-type dyes) when gel analysis is required.

• Energy and cold-chain:

  • Store working aliquots at −20 °C instead of −80 °C when stability permits (this product’s storage is −20 °C per Product Data). Use frost-free practices to improve freezer efficiency.

Tradeoffs: Physical delivery methods may impact cell viability; greener lipids may alter transfection efficiency. Validate performance versus environmental benefits for your specific system.

Pharmaceutical Uses

• Item-specific: For research use only (Product Data). Not for human or veterinary use, diagnostic procedures, or clinical applications.

• General context (non-clinical, informational):

  • siRNA therapeutics in clinical settings require GMP manufacturing, stringent impurity profiles, endotoxin control, and validated delivery systems (e.g., LNPs, conjugates). This product does not claim such status.
  • In drug discovery research, siRNAs are commonly used for target validation, pathway deconvolution, and mechanism-of-action studies in cell-based assays. They can inform whether modulation of a gene like Grm4 yields phenotypes relevant to a discovery program.
  • Formulation roles (research scale): Complexation with cationic lipids/polymers for in vitro delivery; lyophilized formats facilitate storage and shipping. Specific format and excipients for this item are not specified.

Any pharmaceutical or clinical use would require separate, compliant materials and documentation; this catalog item is intended strictly for laboratory research.

Physical Properties

• Item-specific (Product Data):

  • Physical state/appearance: Not specified for this item; refer to CoA/Spec Sheet.
  • Molecular weight and formula: Not specified for this item; refer to CoA/Spec Sheet.
  • Solubility: Supplied as an siRNA reagent set; reconstitution typically in nuclease-free water or buffer (see Storage & Reconstitution). Exact solubility specifications: Not specified for this item; refer to CoA/Spec Sheet.
  • Other specifications (pH, absorbance ratios, OD260): Not specified for this item; refer to CoA/Spec Sheet.

• General properties for siRNA (literature, not item specifications):

  • Typical length: ~19–21 base pairs with 2-nt 3′ overhangs.
  • Charge: Strongly polyanionic due to phosphate backbone; requires cationic carriers for cellular delivery.
  • Solubility: Freely soluble in RNase-free water and low-ionic-strength buffers; avoid divalent cations and RNases.
  • Stability: Dry (lyophilized) siRNA is stable at −20 °C for months to years; in solution, stability improves at ≤−20 °C and with RNase-free conditions; repeated freeze–thaw may degrade RNA.
  • UV detection: Nucleic acids absorb at 260 nm; A260 of 1.0 ≈ 40 μg/mL for single-stranded RNA (approximate literature conversion; duplex siRNA may differ slightly). These are general reference values only.

All authoritative physical/analytical specifications for this specific item should be taken from the CoA/Spec Sheet.

Quality and Grades

• Item-specific: Grade/Purity not specified for this item; refer to CoA/Spec Sheet.

• Context for siRNA quality (general literature/industry practice):

  • Synthesis quality tiers commonly include desalted (research-grade), cartridge-purified, PAGE-purified, or HPLC-purified duplexes. Higher purification reduces truncated species and may improve potency and reduce off-target effects.
  • Identity/quality verification often involves OD260 quantitation, mass spectrometry (MALDI-TOF/ESI), capillary electrophoresis or HPLC profiles, and annealing verification.
  • Modifications (2′-OMe, 2′-F, phosphorothioate, terminal conjugates) can tune nuclease resistance and delivery; if present, they should be documented on the CoA.
  • Endotoxin and bioburden levels can be critical for cell-based assays; acceptance criteria vary by vendor and application. Values for this product are not specified here.

• Practical guidance:

  • For sensitive assays or primary cells, HPLC- or PAGE-purified siRNAs are typically preferred (general guidance).
  • Confirm whether this set includes multiple independent siRNA sequences per target (common for set “A” offerings) and whether they are delivered lyophilized or in solution; details not specified here. Consult the Spec Sheet for sequence list, amount (nmol/duplex), purity, and QC metrics.

Reaction and Applications

• Item-specific: The catalog indicates a “pre-designed siRNA set” for mouse Grm4. Detailed application notes are not provided in the Product Data; refer to the datasheet for assay-validated use cases.

• General applications for siRNA (literature/practice):

  • Gene knockdown in cultured mammalian cells to interrogate gene function, pathway mapping, and target validation.
  • Transient silencing with peak effect typically 24–72 h post-transfection; duration depends on cell division rate and RNA stability.
  • Co-transfection with reporter constructs for pathway readouts; rescue experiments using siRNA-resistant cDNA to confirm on-target effects.
  • Pooling multiple independent siRNAs against the same gene can increase knockdown robustness and reduce off-target effects.

• Practical tips:

  • Optimize siRNA concentration (often 1–50 nM final) and transfection reagent ratio by small-scale matrix experiments.
  • Include controls: non-targeting siRNA, positive control against a housekeeping gene (e.g., PLK1 for cytotoxic readout), and mock-transfection.
  • Assess knockdown by RT-qPCR (mRNA) and by Western blot/ELISA (protein); confirm phenotypes in at least two independent siRNAs.
  • For difficult cells, consider electroporation or alternative delivery systems (lipid nanoparticles, polymer carriers). Primary neurons and immune cells may need specialized protocols.

• Special considerations for GPCR targets (e.g., Grm4; literature):

  • Validate functional impact with second-messenger assays (cAMP), receptor trafficking, or ligand-response profiling, in addition to expression measures.

Reaction Conditions

Classical reaction condition guidance (solvent, temperature, catalysts, yields) is not applicable to siRNA, which functions in biological systems rather than chemical transformations.

• General experimental conditions for siRNA transfection (literature; not item-specific):
  • Complexation: Mix siRNA with a cationic lipid/polymer in serum-free medium (e.g., Opti-MEM) for 5–20 min at room temperature to form nanoparticles.
  • Dosing: Typical final concentrations range 1–50 nM in cell culture; optimal values are cell line and reagent dependent.
  • Incubation: Add complexes to cells at 40–80% confluence; replace with complete medium after 4–24 h depending on transfection reagent guidance.
  • Readouts: Assess mRNA knockdown by RT-qPCR at 24–48 h and protein reduction at 48–96 h.
  • Temperature: Standard mammalian cell culture conditions (37 °C, 5% CO2) unless working with temperature-sensitive cells.

These are general literature practices. For validated, product-specific conditions and controls for the Grm4 Mouse Pre-designed siRNA Set A, consult the product datasheet if available.

Safety and Handling

• GHS/SDS status (item-specific):

  • Signal Word: Not specified for this item; refer to SDS.
  • H-Statements / P-Statements: Not specified for this item; refer to SDS.
  • GHS Classification and Pictograms: Not specified for this item; refer to SDS.

• General safety guidance for siRNA reagents (literature/practice):

  • Expected hazards: Synthetic siRNAs are generally considered low hazard; primary risks are exposure to RNases (loss of activity) and minor irritation from buffers or dust.
  • PPE: Use lab coat, nitrile gloves, and eye protection. Change gloves frequently to minimize RNase contamination.
  • Handling: Work in a clean, RNase-free area. Use certified RNase-free tubes, barrier tips, and dedicated pipettes. Avoid mouth pipetting and aerosol generation.
  • Incompatibilities: Avoid RNase contamination from skin, shared reagents, or non-sterile plasticware. Limit exposure to strong acids/bases and nucleases. Avoid repeated freeze–thaw cycles that can shear RNA.
  • First aid (general): In case of eye/skin contact with solutions, rinse with water. If inhaled (powder/aerosol), move to fresh air. If ingested, rinse mouth. Seek medical advice if symptoms persist. Defer to SDS for authoritative instructions.

• Waste: Dispose of aqueous RNA solutions according to institutional biosafety and chemical waste procedures. Decontaminate surfaces with RNase-inactivating agents or freshly prepared 0.1% DEPC-treated water (where appropriate) followed by autoclaving.

Always consult the product-specific SDS for definitive hazard and handling information.

Solvent Selection

This product is not a solvent; “solvent selection” is interpreted here as choosing appropriate reconstitution and working buffers for siRNA.

• Item-specific:

  • Recommended storage: −20 °C (Product Data). Specific reconstitution buffer and concentration: Not specified for this item; refer to CoA/Spec Sheet.

• General guidance for siRNA (literature/practice):

  • Primary solvent: RNase-free water for initial dissolution; common stock concentrations are 10–100 μM.
  • Buffers: Low-ionic-strength, RNase-free buffers such as 10 mM Tris-HCl, pH 7.4–8.0, with 0.1–1 mM EDTA (TE) can improve long-term stability by chelating divalent cations that catalyze hydrolysis.
  • Avoid: Solutions containing RNases, heavy metal ions, and strong acids/bases. Minimize Mg2+/Ca2+ unless required for specific downstream protocols.
  • Compatibility with transfection: Dilute stocks into serum-free media (e.g., Opti-MEM) for complexation with cationic lipids; follow reagent-specific instructions. After complex formation, many protocols tolerate serum-containing media.
  • Filtration: If sterility is required, 0.22 μm filtration can be used on buffers before mixing with siRNA (filtering duplex RNA itself is typically unnecessary and can lead to loss).

• Comparison (general): RNase-free water maximizes chemical simplicity; TE offers stability; PBS is generally avoided for storage due to ionic strength and divalent cations that may promote degradation.

Storage and Reconstitution

• Item-specific (Product Data):

  • Storage Conditions: Store at −20 °C.
  • Shipping: Shipped in an ice chest with ice pads.

• Additional item-specific details (amount per tube, buffer, concentration, duplex status): Not specified for this item; refer to CoA/Spec Sheet.

• General guidance for siRNA (literature/practice; not item specifications):

  • Upon receipt: Allow the sealed container to equilibrate to room temperature before opening to prevent condensation. Centrifuge briefly to collect contents.
  • Reconstitution: Dissolve in RNase-free water or TE buffer (10 mM Tris-HCl, 0.1–1 mM EDTA, pH 7.4–8.0) to a convenient stock (e.g., 20–100 μM). Mix gently—do not vortex vigorously.
  • Aliquoting: Prepare single-use aliquots to avoid repeated freeze–thaw. Label aliquots with sequence ID and concentration.
  • Storage after reconstitution: Store at −20 °C for medium-term (weeks to months) or at −80 °C for long-term. Protect from repeated freeze–thaw and from RNase contamination. Keep in low-adsorption tubes.
  • Thawing: Thaw on ice, mix by gentle pipetting, and keep on ice during setup. Discard any aliquot suspected of RNase contamination or degradation (smearing on native PAGE or loss of activity).

For any product-specific requirements (e.g., recommended solvent, initial concentration, presence of stabilizers), defer to the product’s CoA/Spec Sheet.

Structure and Identity

This product is a pre-designed small interfering RNA (siRNA) reagent set targeting the mouse Grm4 gene (metabotropic glutamate receptor 4). As a nucleic acid tool, it does not have a conventional small-molecule structure.

• Item-specific (Product Data):

  • SKU: G1469296
  • Product Name: Grm4 Mouse Pre-designed siRNA Set A
  • Type: Pre-designed siRNA duplex set (mouse)
  • CAS: 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.
  • SMILES / InChIKey: Not applicable for short double-stranded RNAs.

• General nucleic acid features (literature):

  • Typical siRNA comprises two ~21-nt strands with 2-nt 3′ overhangs (often dTdT) and a phosphorylated 5′ end on the guide strand.
  • Chemical modifications (e.g., 2′-OMe, 2′-F, phosphorothioate) may be used to enhance stability and reduce off-targeting; presence/absence for this item: Not specified for this item; refer to CoA/Spec Sheet.

• Structural description in words (general):

  • A short, double-stranded RNA helix with antiparallel strands; the guide strand is loaded into RISC to direct sequence-specific mRNA cleavage/silencing. Sequence, length, and modification pattern determine target specificity, potency, and stability. Sequence details for this item are not provided here; consult the product documentation.

Synthetic Utility

This item is a biological knockdown reagent (siRNA) rather than a synthetic building block or reagent; conventional “synthetic utility” in organic synthesis does not apply.

• Not applicable: It is not used to form covalent bonds in chemical synthesis, nor does it serve as a catalyst or stoichiometric reagent in typical organic reactions.

• Relevant utility (general research context):

  • Provides a reversible, sequence-directed means to silence gene expression, enabling functional genomics studies that complement CRISPR/Cas9 knockout or shRNA approaches.
  • Useful in chemical biology to validate on-target effects of small molecules by phenocopying or rescuing phenotypes via gene expression modulation.

For chemical synthesis applications, refer instead to appropriate reagents and catalysts listed elsewhere in the catalog.

Target Specificity

• Item-specific (from Product Name):

  • Target gene: Grm4 (mouse; metabotropic glutamate receptor 4).
  • Product format: Pre-designed siRNA Set A.
  • Number of siRNA duplexes, sequences, and modification status: Not specified for this item; refer to CoA/Spec Sheet.

• Notes on specificity (general literature):

  • Each siRNA within a set typically targets distinct regions of the same transcript to enhance robustness and allow confirmation of on-target effects.
  • Off-target minimization strategies include seed-region optimization and chemical modifications; details for this item are not provided.
  • Validate specificity by demonstrating concordant phenotypes across independent siRNAs and by rescue with siRNA-resistant Grm4 cDNA.

All definitive details on targeting sequences, transcript isoforms, and predicted off-targets should be obtained from the item’s datasheet.