TFF3 Human Pre-designed siRNA Set A

Cat. No.: T1489644

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Panoramica

TFF3 Human Pre-designed siRNA Set A contains three designed siRNAs for TFF3 gene (Human), as well as a negative control, a positive control, and a FAM-labeled negative control. Components TFF3 siRNA-1: 5 nmol (HPLC) TFF3 siRNA-2: 5 nmol (HPLC) TFF3 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

Condizioni di conservazione di stoccaggio

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.

Product Line

Prodotti biologici

Application Protocols

Item-specific validated applications and dilutions: Not specified for this item; refer to CoA/Spec Sheet.

General siRNA transfection workflow (literature/general guidance):

1. Preparation

• Thaw all components on ice. Use RNase-free tubes and tips. If lyophilized, briefly centrifuge and reconstitute siRNA with RNase-free water or TE to a convenient stock (e.g., 10–20 µM).

2. Complex formation

• In serum-free medium (e.g., Opti-MEM), dilute siRNA and transfection reagent separately, then combine and incubate 10–30 minutes at room temperature to allow complexation. Ratios are reagent-specific.

3. Cell exposure

• Add complexes dropwise to cells at the desired confluence. Gently rock to distribute. Maintain per reagent instructions; some protocols allow presence of serum.

4. Post-transfection

• Replace medium if required after 4–24 hours. Assess mRNA knockdown by qRT-PCR at 24–48 h; assess protein levels at 48–96 h.

5. Controls

• Include non-targeting siRNA, positive control siRNA, and mock control. Perform dose titration to optimize potency vs. off-target effects.

6. Replication and validation

• Test individual duplexes from Set A and/or pools. Validate with independent duplexes and, where feasible, rescue by ectopic expression lacking target UTR.

Sterility and RNase control are essential throughout.

Biological Roles

Target gene overview (literature/general knowledge):

• TFF3 (Trefoil Factor 3) encodes a small secreted peptide of the trefoil factor family, predominantly expressed in mucin-secreting epithelial cells of the gastrointestinal and respiratory tracts.
• Molecular function: Trefoil factors are characterized by one or more trefoil domains (three-loop structure stabilized by disulfide bonds) and are implicated in mucosal barrier maintenance, epithelial restitution, and modulation of cell migration under stress conditions.
• Pathways and interactions: TFF3 is co-expressed with gel-forming mucins and can influence epithelial signaling cascades related to restitution and barrier function. It may interact with glycoprotein partners at the mucosal surface; exact receptors remain an area of investigation.
• Research utility: Silencing TFF3 permits interrogation of mucosal defense mechanisms, epithelial wound-healing responses, and adaptive transcriptional programs. In cancer biology research, modulation of TFF3 expression is studied for its effects on cell motility and signaling pathways.

Note: The above reflects general biological knowledge about TFF3. This product is for research use only and is intended to transiently reduce TFF3 mRNA levels via RNA interference to facilitate mechanistic studies. No medical or diagnostic claims are made.

Buffer Applications

Classical buffer formulation guidelines (e.g., buffering capacity, pH curves) are not directly applicable to siRNA as a buffer component. However, siRNA handling relies on appropriate buffers.

General guidance for siRNA buffers (literature/general knowledge):

• Stock preparation: Reconstitute in RNase-free water or TE buffer (10 mM Tris-HCl, 0.1–1 mM EDTA, pH 7.5–8.0). EDTA helps chelate trace divalent cations that can support RNase activity.
• Working solutions: Dilute in serum-free Opti-MEM or equivalent for complex formation with lipid-based transfection reagents, then add to complete media per reagent instructions.
• Ionic strength: Maintain moderate salt conditions for duplex stability; avoid high Mg2+/Ca2+ unless protocol-specific.
• Sterility: For sensitive cell assays, use sterile-filtered buffers and low-binding plastics to minimize adsorption.

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

Green Alternatives

Conventional green chemistry substitution is not directly applicable to siRNA reagents. However, from a sustainability and biosafety standpoint (literature/general knowledge):

• Compared with plasmid/viral shRNA delivery, synthetic siRNA:
  • Avoids antibiotic selection and large-scale bacterial culture (reduced biomass and solvent use).
  • Eliminates viral packaging and associated biohazard levels.
  • Enables transient, titratable knockdown without genomic integration.
• Transfection reagent choice: Consider lower-toxicity, serum-compatible reagents that enable reduced reagent and siRNA doses, decreasing waste and improving cell health.
• Process considerations:
  • Use RNase-free, reusable equipment where possible and certified nuclease-decontamination procedures to minimize consumable waste.
  • Prepare concentrated stocks to reduce plastic usage from multiple dilutions.
• Shipping/storage: Lyophilized formats (if available; item-specific form not specified) can reduce cold-chain energy demands. Consolidate shipments to minimize packaging.

Item-specific environmental/sustainability claims: Not specified for this item; refer to CoA/Spec Sheet.

Pharmaceutical Uses

This product is supplied strictly for research use only. It is not intended for human or animal therapeutic, diagnostic, or clinical applications.

General formulation context for oligonucleotides (literature/general knowledge):

• In pharmaceutical development settings (outside the scope of this research reagent), small interfering RNAs may require specialized delivery systems, isotonic buffers, and stabilizing excipients. Such considerations do not apply to this catalog item in its research-use form.

Item-specific pharmacopeial status, excipient profile, or GMP information: Not specified for this item; refer to CoA/Spec Sheet.

Physical Properties

Item-specific physicochemical specifications (BP, MP, density, UV cutoff, exact solubility): Not specified for this item; refer to CoA/Spec Sheet.

General properties for siRNA oligonucleotides (literature/general knowledge):

• Physical state: Often supplied lyophilized, white to off-white solid, or as an aqueous solution in RNase-free buffer at vendor-specified concentration.
• Solubility: Freely soluble in RNase-free water and common biological buffers (e.g., TE, PBS) to ≥100 µM; avoid divalent cation contamination that may promote nuclease activity.
• UV characteristics: Strong absorbance at 260 nm (A260) with sequence-dependent molar extinction coefficient calculated from nearest-neighbor parameters.
• pKa/logP: Not applicable in conventional sense for polyanionic biopolymers; siRNA is highly hydrophilic and polyanionic at physiological pH due to phosphate groups.
• Stability: Sensitive to RNases; enhanced stability with low temperatures, chelators (e.g., EDTA), and optional backbone/sugar modifications (if present). Item-specific stabilization: Not specified for this item; refer to CoA/Spec Sheet.
• Duplex Tm: Sequence- and salt-dependent; typically 60–80°C for 21–23 bp under moderate salt (literature). Exact Tm for included duplexes: Not specified for this item; refer to CoA/Spec Sheet.

Quality and Grades

Item-specific grade, purity, sequence verification, modification pattern: Not specified for this item; refer to CoA/Spec Sheet.

General notes on siRNA quality (literature/general knowledge):

• Typical grades: Desalted, cartridge-purified, or HPLC/ PAGE-purified. Higher purity (HPLC/PAGE) reduces truncated species and can improve potency and consistency.
• Identity & QC: Vendors commonly provide sequence information, mass confirmation by ESI–MS or MALDI, and OD260-based quantitation. Endotoxin and bioburden testing may be offered for cell-based work.
• Stabilizers: Some siRNAs include chemical modifications (e.g., 2′-O-Me, 2′-F, PS linkages) at select positions to enhance nuclease resistance and reduce immunostimulation. Item-specific stabilizer content: Not specified for this item; refer to CoA/Spec Sheet.
• RNase-free assurance: Manufacturing in RNase-free environments; products are typically supplied in RNase-free vials and may be accompanied by DNase/RNase test results.
• Set design (pre-designed “Set A”): Multiple non-overlapping duplexes targeting human TFF3 mRNA to enable empirical selection of the most effective guide. This strategy increases likelihood of robust knockdown across cell lines and experimental conditions.

Reaction and Applications

This product is intended for gene expression knockdown via RNA interference in human systems targeting TFF3 transcripts. It is not a chemical reagent for traditional organic reactions.

Applications (literature/general knowledge):

• Functional genomics: Transient knockdown of TFF3 to study pathway involvement, compensatory mechanisms, or phenotypic outcomes in epithelial, mucosal, or cancer-derived cell lines.
• Validation workflows: Use multiple duplexes in Set A to identify the most potent and specific guide; confirm knockdown by qRT-PCR (mRNA) and immunoblot/ELISA (protein), followed by rescue experiments where appropriate.
• Assay contexts: Signaling pathway dissection, co-depletion with pathway members, synthetic lethality screens, and transcriptomic profiling (RNA-seq) post-knockdown.

Practical tips:

• Optimize transfection conditions (reagent type, cell density, serum presence, siRNA dose), and include controls (non-targeting siRNA, positive knockdown control, mock).
• Use RNase-free technique and pre-warm complexes per transfection reagent instructions. Titrate siRNA (e.g., 1–50 nM) to balance potency vs. off-target effects; lower doses often improve specificity.
• Pooling strategy: Combining duplexes can improve robustness at lower per-guide concentrations; alternatively, test individually to select a single guide for follow-up.

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

Reaction Conditions

Classical reaction condition guidance (solvent, temperature, catalysts) is not applicable; this product is not used in organic or inorganic synthesis.

Transfection/knockdown conditions (literature/general knowledge):

• Cell density: Optimize per cell line (e.g., 30–70% confluence at transfection).
• Dose: Empirically determine siRNA concentration, often 1–50 nM in culture; lower doses may enhance specificity.
• Complexation: Follow instructions of the chosen transfection reagent for incubation time and ratios; typical room-temperature complexation for 10–30 minutes in serum-free medium.
• Incubation: Replace or supplement medium per reagent guidance; assess mRNA knockdown at ~24–48 h and protein knockdown at ~48–96 h post-transfection.
• Controls: Include non-targeting control, positive control siRNA (e.g., targeting housekeeping gene), and mock transfection.

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

Safety and Handling

GHS/CLP classification, signal word, H-statements, pictograms: Not specified for this item; refer to SDS.

General laboratory safety for siRNA (literature/general guidance):

• Expected hazards: Oligonucleotides are generally regarded as low hazard in standard laboratory use. However, formulation buffers may contain irritants depending on supplier. Always consult the SDS for authoritative guidance.
• PPE: Wear lab coat, gloves (nitrile), and safety glasses. Use RNase-free technique; change gloves frequently to prevent RNase contamination.
• Handling: Work in a clean area dedicated to RNA work. Use RNase-free, nuclease-decontaminated consumables and solutions. Avoid generating aerosols with concentrated stocks.
• Incompatibilities: Avoid RNase contamination (skin, non-sterile plastics), strong oxidizers for certain modified backbones, and repeated freeze–thaw cycles which may degrade RNA.
• First aid (overview; refer to SDS): In case of skin/eye contact with solutions, rinse with water. If inhaled (aerosolized powders), move to fresh air. If ingested, rinse mouth and seek medical attention as needed.
• Waste: Dispose of RNA and consumables as nonhazardous biological lab waste per institutional policy unless mixed with hazardous agents.
• Transport/shipping: Shipped with cold packs as provided. Maintain cold chain for solutions; lyophilized materials are more tolerant but still protect from heat and moisture.

Solvent Selection

For siRNA reagents, classical organic solvent selection is not typically applicable. siRNA is handled in aqueous, RNase-free solutions.

General guidance (literature/general knowledge):

• Primary solvents: RNase-free water, TE buffer (10 mM Tris-HCl, 0.1–1 mM EDTA, pH ~7.5–8.0), or manufacturer-specified buffer. EDTA chelates divalent cations that can support nuclease activity.
• Compatibility: Readily miscible with standard cell culture media during transfection when complexed with a transfection reagent. Avoid high ionic strength or divalent cations during stock preparation.
• Avoid: Organic solvents (e.g., ethanol, DMSO) for stock solutions unless explicitly directed; these can affect duplex integrity and downstream biological assays.
• Working solutions: Prepare at 1–100 nM in culture depending on cell type and reagent; optimize empirically.
• Filtration: If sterility is required, use low protein-binding 0.22 µm filters and minimize shear; however, filtration of very dilute siRNA may lead to loss by adsorption.

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

Storage and Reconstitution

Item-specific storage and shipping:

• Storage conditions: Store at -20°C (as provided in Product Data).
• Shipped in: Ice chest + Ice pads (as provided in Product Data).

General guidance for siRNA (literature/general knowledge):

• Reconstitution: If lyophilized, spin down contents and reconstitute in RNase-free water or TE buffer to a convenient stock concentration (commonly 10–100 µM). Mix gently; avoid vigorous vortexing of very dilute solutions to minimize shear/foaming.
• Aliquoting: Prepare single-use aliquots to avoid repeated freeze–thaw cycles. Use low-binding tubes.
• Storage after reconstitution: Store at -20°C for medium-term (months) or -80°C for long-term storage; avoid frost-free freezers. Protect from light if dye-labeled.
• Freeze–thaw: Minimize cycles; thaw on ice and keep solutions cold during setup.
• Stability: Unmodified siRNA is generally stable for multiple months at -20°C; modified siRNA may exhibit enhanced stability. Exact stability for this item: Not specified for this item; refer to CoA/Spec Sheet.
• Documentation: Record OD260, stock concentration, and lot information upon first use to ensure experimental reproducibility.

Any item-specific buffer composition, concentration on delivery, or stabilizers: Not specified for this item; refer to CoA/Spec Sheet.

Structure and Identity

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

• SKU: T1489644
• Product name: TFF3 Human Pre-designed siRNA Set A
• Target gene: TFF3 (Trefoil Factor 3), species: Homo sapiens (human)
• CAS, CID, InChIKey, SMILES: Not applicable to siRNA mixtures; Not specified for this item; refer to CoA/Spec Sheet.
• Molecular formula and molecular weight: Sequence-dependent; Not specified for this item; refer to CoA/Spec Sheet.

General structural description (literature/general knowledge):

• Pre-designed siRNA sets typically comprise 2–4 distinct double-stranded siRNA duplexes targeting non-overlapping regions of the same mRNA to increase likelihood of potent knockdown while mitigating off-target effects.
• Each siRNA is usually a ~21–23 bp RNA duplex with 3′-dinucleotide overhangs (commonly 3′-UU or 3′-dTdT). Sense strand is complementary to target mRNA; antisense strand (guide) is loaded into RISC.
• Chemical features may include 2′-OH ribose, and, in some formats, stabilizing modifications (e.g., 2′-O-Me, phosphorothioates) at select positions; Item-specific modification status: Not specified for this item; refer to CoA/Spec Sheet.
• Stereochemistry: RNA phosphodiesters are chiral at phosphorus in modified backbones (if present), otherwise canonical RNA phosphodiester linkages.
• Form: Typically supplied lyophilized or in RNase-free solution; Item-specific form: Not specified for this item; refer to CoA/Spec Sheet.

Synthetic Utility

Not typically applicable. This product is a biological reagent (siRNA duplexes) intended for gene knockdown studies rather than as a substrate/reagent in chemical synthesis.

General note (literature/general knowledge):

• Oligoribonucleotides can serve as templates or primers in molecular biology workflows, but they are not used as building blocks in standard organic synthesis or materials chemistry. For nucleic acid chemistry or conjugation studies, custom-modified siRNAs with defined functional handles (e.g., amino, thiol, azide) are typically ordered specifically for that purpose.

Item-specific functional group modifications enabling conjugation: Not specified for this item; refer to CoA/Spec Sheet.

Target Specificity

Item-specific details provided by product name:

• Target: Human TFF3 (Trefoil Factor 3)
• Species reactivity: Human
• Format: Pre-designed siRNA Set A (multiple duplexes targeting TFF3 transcripts)

Additional item-specific details (epitope, clone, isotype—antibody-specific fields): Not applicable to siRNA.

Sequence information, targeted transcript isoforms, off-target analyses: Not specified for this item; refer to CoA/Spec Sheet.

General notes (literature/general knowledge):

• Using multiple non-overlapping siRNAs supports verification of on-target effects. Concordant phenotypes across distinct duplexes increase confidence in target specificity.
• Bioinformatic screening typically minimizes seed-region off-targets and microRNA-like effects; empirical validation (dose-response, rescue) is still essential.