BTBD2 Human Pre-designed siRNA Set A

Cat. No.: B1488599

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Übersicht

BTBD2 Human Pre-designed siRNA Set A contains three designed siRNAs for BTBD2 gene (Human), as well as a negative control, a positive control, and a FAM-labeled negative control. Components BTBD2 siRNA-1: 5 nmol (HPLC) BTBD2 siRNA-2: 5 nmol (HPLC) BTBD2 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.

Application Protocols

Item-specific, tested application details are Not specified for this item; refer to CoA/Spec Sheet.

General workflow outline (informational; not product‑specific)

• Thaw on ice and briefly spin down. Reconstitute each duplex in nuclease‑free water or TE to a convenient stock (e.g., 10–50 µM). Aliquot and store at −20°C.
• Prepare transfection complexes according to the chosen delivery reagent. Optimize siRNA dose (typ. low‑nM) and cell density.
• Include controls: non‑targeting siRNA, positive‑control siRNA, and a mock transfection.
• Assess knockdown at RNA level (qRT‑PCR, 24–48 h) and protein level (immunoblot or imaging, 48–96 h). Validate phenotypes with ≥2 independent duplexes from the set.

Biological Roles

Item-specific facts (from Product Data)

• Target: Human BTBD2 (BTB/POZ domain-containing protein 2)

Literature/general knowledge about BTBD2 (not product‑specific; verify against primary literature for your use case)

• Gene/function: BTBD2 encodes a BTB/POZ domain protein implicated in protein–protein interactions and ubiquitin ligase complexes. Members of the BTB family often act as substrate adaptors for Cullin3 (CUL3) E3 ubiquitin ligases, influencing protein turnover and signaling. Specific roles for BTBD2 have been reported in cytoskeletal regulation and membrane trafficking contexts in several models.
• Cellular localization: Predominantly cytoplasmic with potential association to cytoskeletal or membrane structures reported in some studies.
• Pathways: Putative involvement in ubiquitin‑dependent proteostasis and possibly endocytic/trafficking pathways; detailed mechanisms remain under active investigation.

Use of siRNA against BTBD2 (general guidance)

• Application: Transient knockdown in human cell lines to interrogate BTBD2 function, map interaction partners, and assess phenotypic consequences of reduced expression.
• Controls: Employ non‑targeting siRNA and an siRNA targeting an unrelated gene as negative/orthogonal controls; confirm knockdown by qRT‑PCR and/or immunoblotting with validated BTBD2 antibodies.
• Off‑target minimization: Use pools of independent siRNAs (as provided in pre‑designed sets) and titrate to the minimal effective concentration to reduce seed‑mediated off‑targets.

Buffer Applications

Applicability

• While not a traditional buffer, siRNA handling relies on compatible, RNase‑free buffers to maintain integrity and efficacy.

General guidance (not product‑specific)

• Reconstitution: Use nuclease‑free water or TE buffer (10 mM Tris‑HCl, 0.1–1 mM EDTA, pH 7.5–8.0) to stabilize RNA against trace nucleases and divalent cations. Avoid phosphate buffers for long‑term storage if divalent cations may be present.
• Working solutions: Prepare dilutions in RNase‑free water or Opti‑MEM/reduced‑serum media when complexing with lipid‑based transfection reagents; follow the delivery reagent’s buffer recommendations.
• Electrophoresis checks: For QC, native PAGE or agarose in TBE/TBE‑urea can evaluate duplex integrity; include RNase inhibitor if needed during sample prep.
• Storage buffer ionic strength: Low‑salt buffers reduce aggregation and preserve duplex structure; high salt may affect complexation with cationic lipids.

Item-specific note

• No buffer composition is specified for this product. Refer to the CoA/Spec Sheet for any provided or recommended reconstitution buffer.

Green Alternatives

Relevance

• “Green solvent/reagent” considerations are not directly applicable to siRNA oligonucleotides.

General sustainability considerations (not product‑specific)

• Reduce waste: Prepare aliquots to minimize freeze–thaw waste; scale experiments to the minimum effective volumes.
• Safer auxiliaries: Choose transfection reagents with favorable safety profiles and avoid halogenated solvents in any ancillary steps (e.g., plasmid prep cleanups).
• Alternative modalities: For durable knockdown without repeated reagent use, consider CRISPRi or shRNA approaches (tradeoffs include setup time, vector handling, and biosafety requirements). These are methodological, not “greener” per se.

Conclusion

• Focus on good laboratory practice, right‑sizing experiment scale, and minimizing single‑use plastics to improve sustainability when working with siRNA.

Pharmaceutical Uses

Scope limitation

• For research use only. Not for diagnostic, therapeutic, or in vivo clinical use.

Research/manufacturing context (general; not product‑specific)

• Discovery workflows: Pre‑designed siRNA sets are widely used in target validation, mechanism‑of‑action studies, and phenotypic screens that support early drug discovery decisions.
• Assay development: siRNA knockdown provides orthogonal controls for high‑content imaging, reporter assays, and biochemical readouts to confirm on‑target pharmacology of small‑molecule modulators.
• Formulation parallels: While not an excipient, handling principles align with nucleic acid manufacturing—stringent nuclease control, low bioburden, and validated quantification by A260.

Compliance

• No pharmacopeial monograph or clinical‑grade claims are made for this item. Any GMP, endotoxin, sterility, or residual solvent specifications are Not specified for this item; refer to CoA/Spec Sheet.

Physical Properties

Item-specific facts (from Product Data)

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.

Literature/general values for siRNA (for context; not product specifications)

• Typical duplex length: ~21 bp with 2‑nt 3′ overhangs (commonly dTdT), or 19–27 bp depending on design.
• Approximate MW range: ~13–16 kDa per duplex for 21‑mers (sequence dependent).
• Solubility: siRNA is water‑soluble; commonly reconstituted in nuclease‑free water or TE buffer (10 mM Tris‑HCl, 0.1–1 mM EDTA, pH 7.5–8.0).
• UV characteristics: Strong absorbance at 260 nm (A260); extinction coefficient depends on sequence and length.

Practical notes (general guidance)

• Working concentration range: Typically 1–50 µM stock; final transfection concentrations often 1–50 nM, optimized per cell line and delivery reagent. Not specified for this item.
• pH stability: siRNA is stable near neutral to slightly basic pH in RNase‑free buffers; acidic conditions or divalent RNases can degrade RNA.
• Avoid freeze–thaw cycles; aliquot stocks after initial reconstitution to preserve integrity.

Quality and Grades

Item-specific facts (from Product Data)

• Grade/purity, purification method, stabilizers, endotoxin level: Not specified for this item; refer to CoA/Spec Sheet.

General guidance for siRNA quality (literature/industry practice; not specifications for this item)

• Purification: HPLC‑purified siRNA reduces n‑1 species and truncated products, improving potency and reproducibility. Desalted‑only material may contain more synthesis byproducts and is typically used for screening.
• Identity and integrity: Verified by mass spectrometry and analytical HPLC/UPLC. Duplex formation is confirmed by native PAGE or melting analysis.
• Modifications: Chemical stabilization (e.g., 2′‑O‑Me, 2′‑F, PS linkages) can improve nuclease resistance and in vivo stability; may also reduce innate immune activation. Presence/absence is sequence‑specific—Not specified for this item.
• Low UV background: Not relevant as a solvent grade concept, but OD260 and extinction coefficients are used to quantify siRNA concentration.
• Contaminants: RNase-free status is critical. Endotoxin control can matter for primary cells; typical acceptance criteria vary by application.

Recommendations

• Consult the CoA for sequence(s), OD260 units, μmol scale, purification level, and measured purity. For demanding applications (e.g., primary cells), verify endotoxin and residual salt levels and consider HPLC‑purified, duplex‑annealed formats.

Reaction and Applications

Not typically applicable as a chemical reaction reagent.

Context for use (general, not product‑specific)

• Functional application: Gene expression knockdown via RNA interference (RNAi). The antisense siRNA strand guides RISC to complementary BTBD2 mRNA, promoting Argonaute‑mediated cleavage and reduced protein expression.
• Experimental areas: Target validation, pathway dissection, phenotypic screening, genetic interaction studies, and controls for CRISPR loss‑of‑function.
• Practical tips: Maintain RNase‑free technique; optimize transfection conditions (cell density, reagent choice, siRNA dose). Use multiple independent siRNA duplexes and include non‑targeting and positive‑control siRNAs to confirm on‑target effects.

For step‑by‑step execution, see Application Protocols; item‑specific tested protocols are Not specified for this item; refer to CoA/Spec Sheet.

Reaction Conditions

Not applicable as a chemical reaction component.

General experimental conditions for siRNA transfection (informational only; not product‑specific)

• Temperature: Standard mammalian cell culture conditions (37°C, 5% CO2) following complexation at room temperature per transfection reagent instructions.
• Time: Knockdown kinetics commonly assessed 24–96 h post‑transfection, depending on mRNA/protein half‑life.
• Media: Serum‑free or reduced‑serum media during complex formation; complete media for culture maintenance after addition.

Item-specific, validated reaction/assay conditions are Not specified for this item; refer to CoA/Spec Sheet and the SDS of any ancillary transfection reagent.

Safety and Handling

Item-specific facts (from Product Data)

• GHS classification, pictograms, signal word, H‑statements: Not specified for this item; refer to SDS.
• Storage conditions: Store at −20°C.
• Shipping: Shipped in ice chest + ice pads.
• Research use note: For research use only.

General safety guidance for siRNA and oligonucleotides (not product‑specific)

• PPE: Laboratory coat, gloves, and safety glasses recommended. Handle using RNase‑free technique to prevent sample degradation.
• Incompatibilities: Avoid RNase contamination (skin contact, non‑dedicated plastics, non‑filtered tips). Strong acids/bases and nucleases can degrade RNA. Do not mix with oxidizing sterilants (e.g., sodium hypochlorite + acids) that can generate chlorine.
• Hygiene: Work in a clean area; use RNase‑free water, certified RNase‑free tubes and tips, and clean benches with RNase decontamination solutions.
• First aid (general): In case of contact with eyes/skin, rinse with water. If inhaled (powder/aerosol), move to fresh air. If ingested, rinse mouth with water. Seek medical advice if any symptoms persist. Defer to SDS for authoritative instructions.
• Waste: Dispose of solutions and consumables as non‑hazardous laboratory biological/chemical waste per institutional procedures unless local rules specify otherwise.

Special risks

• Nucleic acids are generally low hazard; primary risks are ergonomic/biological contamination and potential effects from transfection reagents. Review the SDS of any delivery reagent used in conjunction with this product.

Solvent Selection

Applicability

• This product is an aqueous, RNase‑sensitive oligonucleotide reagent rather than an organic solvent. Classical solvent selection concepts (polarity parameters, dielectric constant tables) do not apply.

Practical guidance (general; not product‑specific)

• Reconstitution solvents: Use nuclease‑free water or TE buffer (10 mM Tris‑HCl, 0.1–1 mM EDTA, pH 7.5–8.0). Avoid Mg2+/Ca2+ in storage buffers to limit nuclease activity.
• Working buffers: For cell transfection, dilute into serum‑free or reduced‑serum media when forming complexes with lipid‑based reagents; then add to complete media per the transfection reagent instructions.
• Avoid organic solvents (e.g., ethanol, DMSO) for stock solutions; they can impair duplex structure and/or transfection complexation unless explicitly required by a specific protocol.

Storage and Reconstitution

Item-specific facts (from Product Data)

• Storage: Store at −20°C.
• Shipping: Ice chest + ice pads.

General guidance (not product‑specific)

• Upon receipt: If lyophilized, allow to equilibrate to room temperature in a desiccated environment before opening to prevent condensation.
• Reconstitution: Dissolve in nuclease‑free water or TE buffer (10 mM Tris‑HCl, 0.1–1 mM EDTA, pH 7.5–8.0) to prepare a concentrated stock (e.g., 10–50 µM). Mix gently; avoid vigorous vortexing that may shear nucleic acids.
• Aliquoting: Divide into single‑use aliquots to avoid freeze–thaw cycles. Label with concentration (μM), date, and duplex ID.
• Storage conditions: −20°C for medium‑term storage; for long‑term storage, −80°C is commonly used. Protect from repeated freeze–thaw and from RNase contamination. Avoid frost‑free freezers with temperature cycling.
• Working stability: Thawed aliquots kept on ice during setup are typically stable for the day; discard any leftover diluted working solutions to minimize RNase exposure.
• Quantification: Determine concentration by A260 using the manufacturer‑supplied extinction coefficient or sequence‑calculated values (if provided). For this item, specific coefficients are Not specified; refer to CoA/Spec Sheet.

Structure and Identity

Item-specific facts (from Product Data)

• Product type: Pre-designed small interfering RNA (siRNA) set targeting human BTBD2
• SKU: B1488599
• Product Name: BTBD2 Human Pre-designed siRNA Set A
• 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 / InChI / InChIKey: Not applicable for oligonucleotide mixtures; not specified for this item.

General description (literature/general knowledge)

• Composition: siRNA sets typically comprise 2–4 double‑stranded RNA duplexes (each ~19–27 bp) with 2-nt 3′ overhangs, designed to independently target the same mRNA to increase the probability of efficient knockdown while minimizing off‑target risk.
• Structural features: Each duplex comprises an antisense (guide) strand complementary to the BTBD2 mRNA target region and a sense (passenger) strand. Chemical modifications (e.g., 2′‑O‑methyl, phosphorothioate linkages) may be used for stability in some products; Not specified for this item; refer to CoA/Spec Sheet.
• 2D description in words: Two antiparallel RNA strands forming an A‑form duplex with Watson–Crick base pairing and short 3′ overhangs; terminal phosphate/hydroxyl states and modification patterns depend on synthesis and purification.

Notes

• Because this is a set of oligonucleotide duplexes, a single molecular identity (SMILES, InChI) is not meaningful. Sequence‑level information and modification details should be taken from the Certificate of Analysis (CoA).

Synthetic Utility

Not applicable.

• This is a biological oligonucleotide reagent (siRNA), not a small‑molecule synthetic building block. It is not typically used as a reactant/intermediate in organic synthesis. For relevant usage, see Biological Roles, Buffer Applications, and Application Protocols (where available).

Target Specificity

Item-specific facts (from Product Data only)

• Target gene: BTBD2
• Species reactivity: Human
• Format: Pre‑designed siRNA Set A (multiple duplexes targeting BTBD2)

Notes

• Sequence(s), target transcript regions, and any chemical modifications are Not specified for this item; refer to CoA/Spec Sheet.
• Clone/isotype fields are not applicable to siRNA reagents.