ARFGAP3 Human Pre-designed siRNA Set A

Cat. No.: A1479678

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Übersicht

ARFGAP3 Human Pre-designed siRNA Set A contains three designed siRNAs for ARFGAP3 gene (Human), as well as a negative control, a positive control, and a FAM-labeled negative control. Components ARFGAP3 siRNA-1: 5 nmol (HPLC) ARFGAP3 siRNA-2: 5 nmol (HPLC) ARFGAP3 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

Biologische Produkte

Application Protocols

• Tested applications, recommended dilutions/concentrations, and validated cell lines/conditions are Not specified for this item; refer to CoA/Spec Sheet or the product’s technical datasheet.
• As per developer instructions, only product data should populate this section; no additional protocols are provided here.

Biological Roles

Item-specific statement

• This product is designed for RNAi-mediated silencing of human ARFGAP3 mRNA (gene target inferred from product name). Specific sequences, transcript isoforms, and exon targeting regions are Not specified for this item; refer to CoA/Spec Sheet.

Literature overview of ARFGAP3 biology (not item-specific)

• ARFGAP3 (ADP-ribosylation factor GTPase-activating protein 3) is a member of the ARF-GAP family that inactivates ARF small GTPases by accelerating GTP hydrolysis, modulating vesicle formation and trafficking.
• Cellular localization: Predominantly Golgi-associated; implicated in retrograde transport between Golgi and ER and in endosomal sorting.
• Functional domains: Contains a conserved zinc-finger ARFGAP domain essential for GAP activity; additional regions may mediate membrane association and protein–protein interactions.
• Biological processes: Regulates coat protein complex (e.g., COPI)-dependent trafficking, Golgi integrity, and cargo sorting; contributes to cellular secretion and membrane dynamics.
• Research relevance: Knockdown enables assessment of ARF signaling pathways, Golgi morphology/function, and vesicular transport kinetics; useful for dissecting redundancy/compensation within ARFGAP family members.

Note: Always verify expression in your model system (cell line/tissue) and confirm knockdown at mRNA/protein levels for robust interpretation.

Buffer Applications

Item-specific buffer guidance

• Not specified for this item; refer to CoA/Spec Sheet for any provided buffer or recommended diluent.

General/literature guidance for siRNA handling buffers

• Reconstitution: RNase-free water is recommended initially; optional dilution into 1× TE (10 mM Tris-HCl, 1 mM EDTA, pH 7.5–8.0) for storage stability.
• Working solutions: Prepare in sterile, RNase-free water or buffer compatible with the chosen transfection reagent (often serum-free medium such as Opti-MEM for complex formation).
• pH and ionic strength: Maintain near-neutral pH (7.0–8.0); avoid high salt and divalent cations that can promote degradation or interfere with transfection complexation.
• Aliquoting: Prepare single-use aliquots to avoid freeze–thaw–induced hydrolysis; typical working concentrations are 1–10 µM.
• Quantification: Use A260 to quantify; calculate concentration using extinction coefficients provided by the vendor (Not specified for this item; refer to CoA/Spec Sheet).

Electrophoresis/checks

• Native PAGE or agarose with appropriate dyes can verify duplex integrity; avoid ethidium bromide exposure where possible; SYBR-safe alternatives are preferred.

Green Alternatives

Context

• siRNA workflows are inherently aqueous and typically align with green chemistry principles compared to plasmid/viral methods that may require larger reagent volumes and generate plastic/biological waste.

Greener practice suggestions (literature/general)

• Prefer aqueous buffers (RNase-free water, TE) over organic co-solvents; avoid phenol/chloroform nucleic acid extractions unless necessary.
• Use lipid transfection reagents with lower cytotoxicity and reduced solvent content; consider biodegradable or peptide-based delivery systems to minimize persistent waste.
• Right-size experiments to reduce consumable waste; use multiwell formats and automation to minimize pipette tip and plate usage.
• Implement RNase-free, reusable glassware where appropriate and validated.

Comparison (general; not item-specific)

• Approach: siRNA transfection vs. plasmid-based shRNA
  • Solvents: Aqueous vs. bacterial culture media/antibiotics and DNA miniprep solvents
  • Hazard profile: Minimal chemical hazards vs. potential use of ethidium bromide/organic solvents in cloning workflows
  • Waste: Less biohazardous DNA/vector waste; reduced antibiotic resistance concerns

Trade-offs

• siRNA provides transient knockdown (greener, simpler) but may require repeated dosing; viral/shRNA offers stable expression but with higher biosafety and waste considerations.

Pharmaceutical Uses

• This product is provided strictly for research use only, as stated in the Product Data. It is not intended for human or veterinary use, diagnostic procedures, or as an active pharmaceutical ingredient (API).
• No pharmacopeial status, excipient role, or GMP manufacturing details are provided for this item. Any formulation, clinical, or therapeutic applications are outside the scope of this product.
• For investigators developing therapeutic oligonucleotides, separate GMP-grade materials, endotoxin specifications, sterile filtration/aseptic processing, and regulatory documentation would be required; such attributes are Not specified for this item.

Physical Properties

Item-specific physical specifications

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
• Solubility: siRNA is water-soluble in RNase-free water or buffer; exact solubility for this item Not specified; refer to CoA/Spec Sheet.

Literature/general values for siRNA (for context; not item-specific specs)

• State: solid (lyophilized) or aqueous solution, depending on packaging.
• Solubility: readily soluble in RNase-free water and low-ionic-strength buffers (e.g., 10 mM Tris-HCl, pH 7.5–8.0) to ≥100 µM.
• UV absorbance: λmax ~260 nm; A260 of 1.0 corresponds to ~33 µg/mL for single-stranded RNA; duplex extinction is sequence-dependent.
• pKa/logP: not applicable to polyanionic biopolymers; behavior dominated by phosphate diester acidity and hydration.
• Stability: duplexes are stable at −20 °C; susceptible to RNases in aqueous environments; stabilization by low ionic strength, pH ~7–8, and Mg2+-free buffers.

Note: Do not treat literature values as specifications. Always consult the item’s CoA/SDS for definitive properties.

Quality and Grades

Item-specific quality information

• Grade/purity: Not specified for this item; refer to CoA/Spec Sheet.
• Set composition: “Pre-designed siRNA Set A” typically denotes multiple, independently validated siRNA duplexes targeting distinct regions of the human ARFGAP3 transcript; exact number, sequences, and modification patterns are Not specified for this item; refer to CoA/Spec Sheet.

General notes on siRNA quality (literature/industry practice)

• Purification: Research-grade siRNAs are commonly synthesized by solid-phase phosphoramidite chemistry and purified (e.g., HPLC or cartridge). Higher-purity (HPLC/dual-HPLC) reduces truncated species and improves knockdown consistency.
• Identity/QC: Vendor CoA typically includes OD260, nmol yield, duplex formation/annealing status, and, when applicable, MS or LC traces. Endotoxin testing may be provided for cell-based assays.
• Stabilizers/modifications: Chemical modifications (2′-OMe, 2′-F, PS linkages) can enhance nuclease resistance and reduce innate immune activation; presence/extent for this item is Not specified for this item; refer to CoA/Spec Sheet.
• UV/background: Low UV-absorbing buffers (e.g., nuclease-free water or 1× TE) are preferred for accurate A260 quantification.

How to use grade information in selection

• Choose higher-purity, modification-stabilized siRNA for difficult-to-transfect cells or longer experiments. For routine screening, standard research-grade is often sufficient.

Reaction and Applications

Item-specific applications from catalog

• Manufacturer Applications: Not specified.

General/literature applications for pre-designed siRNA targeting ARFGAP3 (research context)

• Gene silencing: Sequence-specific knockdown of human ARFGAP3 mRNA via RISC-mediated cleavage, enabling transient loss-of-function studies in mammalian cells.
• Pathway dissection: ARFGAP3 encodes an Arf GTPase-activating protein implicated in vesicular trafficking and Golgi-ER dynamics; silencing supports studies of membrane trafficking, secretion, and ARF signaling.
• Methodological uses: Optimize transfection conditions, dose–response, and off-target controls (scrambled/non-targeting siRNA; rescue with siRNA-resistant cDNA) for rigorous phenotypic attribution.
• Multi-duplex strategy: Pre-designed “Set A” typically includes multiple siRNAs to mitigate off-target effects and sequence-dependent variability; convergent phenotypes across duplexes strengthen conclusions.

Practical notes

• Validate knockdown by RT-qPCR (mRNA) and immunoblotting (protein) at 24–72 h post-transfection.
• Include cytotoxicity readouts (e.g., CellTiter-Glo) to separate on-target effects from viability artifacts.
• Consider chemical modifications to enhance stability and reduce immunostimulation in primary cells; presence of such modifications for this item is Not specified for this item; refer to CoA/Spec Sheet.

Reaction Conditions

Item-specific conditions

• Not specified for this item; consult the CoA/Spec Sheet and SDS for any provided guidance.

General/literature conditions for siRNA transfection and knockdown (research use)

• Cell density: Plate adherent cells at ~30–60% confluence on transfection day to allow growth during knockdown window.
• Dose: Common starting range is 5–50 nM total siRNA; optimize per cell type and duplex.
• Complexation: Prepare siRNA–transfection reagent complexes in serum-free medium (e.g., Opti-MEM) for 5–20 min at room temperature per reagent instructions.
• Exposure: Add complexes dropwise; after 4–6 h, replace with complete medium if required by reagent.
• Incubation: Assess mRNA knockdown at 24–48 h; protein knockdown at 48–96 h depending on protein half-life.
• Controls: Include non-targeting siRNA, positive control siRNA (to a housekeeping gene), and mock-transfection controls; perform dose–response to minimize off-target/cytotoxic effects.
• Readouts: RT-qPCR for transcript levels; Western blot/ELISA or phenotypic assays for functional validation.

Notes

• Primary cells and difficult lines may require lower doses and/or chemically modified siRNA; modification status for this item is Not specified for this item.
• For pooled sets, maintain equal molar contribution of each duplex to the total siRNA concentration.

Safety and Handling

Item-specific hazard information

• GHS classification, signal word, pictograms, H-statements: Not specified for this item; refer to SDS.
• Research Use: For research use only (not for diagnostic or therapeutic use).

General safety guidance for RNA oligonucleotides (literature/standard practice)

• Expected hazards: Non-volatile, non-combustible; low acute toxicity anticipated. Primary risks are associated with contamination (RNases) and transfection reagents used in downstream applications.
• PPE: Laboratory coat, gloves (nitrile), and safety glasses. Use RNase-free techniques (RNase-free consumables, fresh gloves) to prevent degradation.
• Handling: Work on clean, RNase-decontaminated surfaces. Avoid repeated freeze–thaw cycles; aliquot upon first thaw.
• Incompatibilities: Avoid strong acids/bases (can hydrolyze RNA) and nuclease contamination. Do not mix with bleach when guanidinium salts (from RNA workstations) are present.
• First aid (overview; defer to SDS): If inhaled or ingested, move to fresh air/rinse mouth with water; seek medical attention if symptoms persist. On skin/eye contact, rinse with water for several minutes.
• Waste: Dispose according to institutional guidelines for non-hazardous biological reagents; follow local regulations, especially if mixed with transfection agents or cytotoxic compounds.

Always consult the SDS for authoritative safety and hazard information.

Solvent Selection

Applicability to this compound type

• As an RNA duplex, this product is handled exclusively in aqueous, RNase-free media; organic solvent selection is generally not applicable.

Aqueous media guidance (literature/general)

• Primary diluent: RNase-free water (nuclease-free) for initial reconstitution.
• Storage buffer: 1× TE (10 mM Tris-HCl, 1 mM EDTA, pH 7.5–8.0) helps chelate divalent cations that catalyze RNA hydrolysis.
• Working buffer: Opti-MEM or serum-free, antibiotic-free medium during transfection; return to complete medium after complex formation and application.
• Ionic strength and divalent cations: Avoid Mg2+/Mn2+ unless protocol requires; divalents can promote RNase activity and backbone cleavage.

Miscibility/polarity context

• siRNA is highly polar and polyanionic; insoluble in nonpolar/less polar organic solvents. Short exposures to alcohols (e.g., ethanol precipitation) are used for purification but not routine handling.

Practical tips

• Use only RNase-free, low-retention plasticware.
• Filter-sterilize buffers (0.22 µm) when sterile conditions are required.
• Avoid repeated freeze–thaw by preparing single-use aliquots at 10–50 µM.

Storage and Reconstitution

Item-specific instructions (from Product Data)

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

General/literature best practices (not item-specific; follow if consistent with above)

• Upon receipt: If lyophilized, allow the vial to equilibrate to room temperature in a desiccated environment before opening to prevent moisture condensation.
• Reconstitution: Dissolve in RNase-free water to a convenient stock (e.g., 50–100 µM); mix gently without vortexing to minimize shearing and foaming.
• Buffering for stability: Optional dilution into 1× TE (10 mM Tris-HCl, 1 mM EDTA, pH 7.5–8.0) for medium-term storage; EDTA chelates divalent cations that promote hydrolysis.
• Aliquoting: Prepare single-use aliquots; avoid repeated freeze–thaw cycles.
• Storage after reconstitution: −20 °C (short-to-medium term) or −80 °C (long term) in RNase-free tubes; protect from light and nuclease contamination.
• Thawing: Thaw on ice, flick to mix, quick spin to collect contents; keep on ice during setup.
• Concentration determination: Measure A260; use extinction coefficients provided by the supplier (Not specified for this item; refer to CoA/Spec Sheet).

Note: If the product is supplied pre-annealed as a duplex vs. single strands, follow any vendor-specific annealing instructions provided in the CoA/Spec Sheet (not specified here).

Structure and Identity

Item-specific facts (from Product Data)

• SKU: A1479678
• Product name: ARFGAP3 Human Pre-designed siRNA Set A
• Intended target: human ARFGAP3 mRNA (gene symbol: ARFGAP3)
• CAS, CID, InChIKey, SMILES: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular formula and molecular weight: Not specified for this item; refer to CoA/Spec Sheet.

General/literature description of siRNA structure (not item-specific)

• Short interfering RNA (siRNA) reagents are typically 19–27 nt antisense/sense strands forming a ~21 bp duplex with 2-nt 3′ overhangs (e.g., dTdT).
• Chemical features may include terminal phosphates, 2′-OMe/2′-F ribose or phosphorothioate linkages for stability; presence and extent of such modifications for this item are Not specified for this item; refer to CoA/Spec Sheet.
• 2D structural description: two antiparallel RNA strands (5′→3′ sense and 5′→3′ antisense) paired by Watson–Crick base pairing, typically with 5′-phosphate/3′-hydroxyl ends.
• Typical duplex MW: ~13–15 kDa per 21-mer duplex (literature, sequence-dependent).

Synthetic Utility

Not typically applicable.

• As an siRNA duplex intended for gene silencing in biological systems, this product is not used as a synthetic reagent, catalyst, or building block in chemical synthesis.
• For nucleic acid chemistry method development, generic siRNA/oligonucleotides may serve as test substrates for ligation, conjugation, or modification workflows; however, sequence-specific ARFGAP3-directed siRNA sets are primarily functional genomics tools rather than synthetic intermediates.

Target Specificity

Item-specific information

• Intended target: human ARFGAP3 mRNA (inferred from product name “ARFGAP3 Human Pre-designed siRNA Set A”).
• Species reactivity: Human (by design intent); cross-reactivity with other species is not specified and depends on sequence homology.
• Duplex sequences, target exons/isoforms, off-target assessment, and validation data: Not specified for this item; refer to CoA/Spec Sheet.

Guidance (general)

• Verify target transcript accession(s) and isoform coverage relevant to your model.
• Perform BLAST or vendor-provided off-target analysis to assess potential cross-reactivity.
• Confirm knockdown with at least two independent siRNA duplexes from the set to strengthen on-target attribution.

Häufig gestellte Fragen

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.