UQCRB Human Pre-designed siRNA Set A

Cat. No.: U1479554

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Panoramica

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

No item-specific, tested application protocols (e.g., recommended transfection reagent, dilution, or validated cell lines) are provided in the Product Data.

Please refer to:

• The product’s CoA/Spec Sheet for any lot-specific handling instructions if available.
• Your chosen transfection reagent’s protocol for complex formation, dosing, and media changes.
• Standard RNAi guidelines in the literature for control selection, endpoint timing, and data analysis.

Research use only.

Biological Roles

Target gene overview (UQCRB; general biology):

• UQCRB encodes the ubiquinol–cytochrome c reductase binding protein, an integral component of mitochondrial respiratory Complex III (cytochrome bc1 complex). It participates in electron transfer from ubiquinol to cytochrome c via the Q-cycle, contributing to proton translocation and maintenance of the proton-motive force across the inner mitochondrial membrane.
• Functional context: Proper UQCRB activity supports oxidative phosphorylation (OXPHOS), ATP synthesis, and influences mitochondrial membrane potential and cellular redox balance. Perturbation can alter ROS generation and downstream signaling related to metabolism and stress responses.
• Cellular localization: Mitochondrial inner membrane; associates with other Complex III subunits within respiratory supercomplexes (literature).

Research implications (general):

• Studying UQCRB knockdown enables dissection of Complex III function, bioenergetics, and adaptive responses such as metabolic rewiring or activation of antioxidant pathways.
• Systems-level readouts include oxygen consumption rate (OCR), extracellular acidification rate (ECAR), metabolite profiling, and proteomic changes in mitochondrial pathways.

Note: The above describes biological roles of the target gene/protein and does not imply any therapeutic or clinical application. No item-specific functional validation data are provided for this siRNA set.

Buffer Applications

While siRNA is not used as a buffer component, proper buffer selection is critical for reconstitution, storage, and transfection.

General buffer guidance (literature):

• Stock reconstitution: RNase-free water or TE buffer (e.g., 10 mM Tris-HCl, 0.1–1 mM EDTA, pH 7.5–8.0) to 10–100 µM. TE can enhance stability for freezer storage by chelating Mg2+.
• Working dilutions: Prepare in nuclease-free water or serum-free medium immediately before complexing with transfection reagent. Final siRNA concentration in cells often 5–50 nM, optimized per cell line and reagent.
• Salt and divalent cations: Keep low during complex formation with cationic lipids/polymers to favor complex stability; switch to complete medium after complexing per reagent instructions.
• Filtration: If sterile solutions are required, use 0.22 µm low-protein-binding filters; avoid shear from repeated plunging through narrow-gauge needles.

Stability considerations:

• Avoid repeated freeze–thaw; aliquot stocks.
• Maintain near-neutral pH; avoid acidic buffers that accelerate RNA hydrolysis.

Item-specific buffer specifications, concentrations, and reconstitution volumes: Not specified for this item; refer to CoA/Spec Sheet.

Green Alternatives

Environmental considerations for siRNA workflows are centered on minimizing hazardous reagents and plastic waste rather than swapping reaction solvents.

• Greener choices (general):
  • Use aqueous buffers (RNase-free water/TE) and avoid unnecessary organic co-solvents.
  • Select transfection reagents with lower cytotoxicity and reduced hazard classifications when performance allows.
  • Scale experiments efficiently and consolidate plates to reduce consumables.
  • Implement nuclease decontamination with less hazardous agents (e.g., certified non-corrosive RNase decontaminants) in place of high-bleach usage where allowed.

Comparison (general):

• Traditional cationic lipids: High efficiency but may carry aquatic toxicity concerns; require careful waste handling.
• Newer polymeric or lipid-like carriers: Some offer similar performance with improved biocompatibility; verify with vendor SDS and literature.

Waste and energy:

• Store at -20 °C as specified; minimize freeze–thaw by aliquoting to reduce product loss and repeated freezer door openings.
• Deactivate nucleic acids in waste only as required by institutional policy; avoid excessive bleach which increases chloride load.

Note: There are no item-specific green certifications or solvent alternatives specified for this product.

Pharmaceutical Uses

This product is designated For research use only. It is not intended for use in humans, diagnostic procedures, or as a therapeutic.

Formulation/excipient context:

• No pharmacopeial grade, excipient role, or GMP status is provided for this item.
• siRNA products for pharmaceutical development require specialized chemistries (e.g., extensive backbone/sugar modifications, conjugates) and stringent CMC controls; such attributes are not specified for this research reagent.

Item-specific regulatory/compendial status: Not specified for this item; refer to CoA/Spec Sheet if available.

Physical Properties

Item-specific physical specs are sequence- and formulation-dependent for siRNA and are not provided here.

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular Weight: Not specified for this item; refer to CoA/Spec Sheet (each duplex MW depends on sequence and any modifications).
• Molecular Formula: Not specified for this item; refer to CoA/Spec Sheet.
• BP/MP: Not applicable for oligonucleotide duplexes (decompose before boiling; melt refers to duplex denaturation, not phase change).
• Solubility (general, literature): Readily soluble in nuclease-free water or buffer (e.g., 10 mM Tris, 0.1–1 mM EDTA, pH 7–8). Avoid divalent cations without chelators to minimize nuclease activity.
• pKa/logP: Not meaningful for large polyanionic biopolymers; siRNA behaves as a strongly anionic polyelectrolyte.
• UV properties (general): Strong absorbance at 260 nm; A260 of 1.0 corresponds roughly to ~40 µg/mL for single-stranded RNA (literature), but exact conversion for duplexes varies with sequence; do not use as item-specific spec.
• Duplex thermal behavior (literature): siRNA exhibits sequence- and salt-dependent melting temperature (Tm), often 55–80 °C in physiological salt; actual Tm must be measured or calculated per sequence.

Note: For exact concentrations, extinction coefficients, and reconstitution volumes, consult the item’s CoA/Spec Sheet if provided.

Quality and Grades

• Grade/Purity: Not specified for this item; refer to CoA/Spec Sheet.

Context for siRNA quality (general guidance):

• Desalting vs. HPLC: Research-use siRNAs are commonly supplied desalted or HPLC-purified. HPLC purification reduces truncated sequences and impurities, often improving knockdown consistency, especially at lower doses. The purification level is not specified for this item.
• Duplexing: Pre-annealed duplexes are typically delivered ready-to-reconstitute; strand stoichiometry is critical for efficacy. The annealing status is not specified for this item.
• End modifications and stabilizers: Optional 3' dTdT overhangs, 2'-O-methyl or phosphorothioate linkages can enhance stability/off-target profile. Presence/absence of modifications is not specified for this item.
• Pre-designed Set A: Generally indicates multiple distinct siRNA sequences against UQCRB to mitigate sequence-specific variability and off-target effects. Exact sequence count and design rules (e.g., seed GC content, off-target screens) are not specified for this item.
• QC analytics (typical, literature): Mass spectrometry and analytical HPLC are commonly used to confirm identity and purity; UV A260 used for quantitation. Item-specific QC methods/results are not specified.

Recommendation: Consult the CoA/Spec Sheet for exact purification method, amount (nmol/µg), and analytical data to plan dosing and assay timelines.

Reaction and Applications

This product is intended for RNA interference (RNAi)-mediated, sequence-specific knockdown of human UQCRB expression in cultured cells for research purposes.

Typical research workflows (general, literature):

• Transient gene silencing: Deliver siRNA via lipofection or polymer-based transfection into human cell lines; assess mRNA reduction by RT-qPCR and protein depletion by Western blot or immunoassay 24–96 h post-transfection.
• Phenotypic screening: Use the multi-duplex set to validate on-target effects—consistent phenotypes across distinct sequences strengthen target attribution.
• Pathway analysis: UQCRB encodes a Complex III subunit; knockdown can perturb electron transport, oxygen consumption rates (OCR), and reactive oxygen species (ROS) profiles. Measure mitochondrial function via Seahorse assays, JC-1/TMRE dye loading, or high-resolution respirometry (general guidance).
• Rescue experiments: Co-transfect siRNA-resistant UQCRB cDNA (silent mutations in the seed region) to confirm specificity (standard RNAi practice).

Practical tips (general):

• Begin with a dose range (e.g., 5–50 nM total siRNA) to optimize viability vs. knockdown; final conditions depend on cell type and transfection reagent.
• Use nontargeting siRNA and positive-control siRNA (e.g., targeting a housekeeping gene) as controls.
• Pooling vs. individual duplexes: Pooled delivery can increase robustness; testing individually allows sequence ranking and off-target evaluation.

Note: No item-specific sequences or validation data are provided; consult the CoA/Spec Sheet if available.

Reaction Conditions

Applicable here as transfection/knockdown conditions (general guidance from literature). Item-specific conditions are not provided.

• Cell types: Optimize per line; adherent mammalian cells commonly respond well to lipid-based transfection.
• Dose: Typical starting screen 5–50 nM total siRNA (e.g., 10 nM, 25 nM, 50 nM) to balance efficacy and viability.
• Complex formation: Mix siRNA with transfection reagent in serum-free medium (e.g., Opti-MEM) for 5–20 minutes at room temperature per reagent protocol.
• Plating density: 30–70% confluence at transfection often yields best uptake; adjust for growth rate.
• Incubation: 4–6 h in complexing medium, then replace with complete growth medium (or as recommended by the reagent vendor). Some reagents allow direct addition to complete medium.
• Readouts: mRNA reduction detectable by 24–48 h; protein reduction typically 48–96 h depending on turnover.
• Repeat dosing: For long experiments (>4–5 days), re-dose at 72–96 h if viability permits.
• Controls: Non-targeting siRNA, positive control siRNA, and mock-transfection are essential for interpreting phenotypes.
• Temperature: Standard cell culture conditions (37 °C, 5% CO2) unless working with specialized systems.

Note: Exact timing, ratios (µL reagent per pmol siRNA), and plate formats should follow the chosen transfection reagent’s instructions. Item-specific optimization data are not specified for this product.

Safety and Handling

GHS/SDS information:

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

General laboratory safety (best practice):

• Wear appropriate PPE: lab coat, gloves, and safety glasses.
• Practice RNase-free technique: use RNase-free tips/tubes, dedicated reagents, and clean benches with RNase decontamination solutions to prevent degradation of siRNA.
• Avoid repetitive freeze–thaw cycles; aliquot upon first thaw to maintain integrity.
• Incompatibilities: Avoid strong acids/bases and nucleases; keep away from DNase/RNase-contaminated solutions and surfaces.
• Transfection operations: Follow the transfection reagent SDS. Some cationic lipids/polymers can be irritants or harmful; handle in a biosafety cabinet when working with cells.

First aid (overview; defer to SDS):

• Skin/eye contact with solutions: Rinse with water for several minutes; remove contaminated clothing; seek medical attention if irritation persists.
• Inhalation/ingestion: Unlikely under normal handling; seek medical attention if exposure occurs.

Waste disposal:

• Dispose of nucleic acid solutions and contaminated consumables according to institutional biosafety and chemical waste procedures. Deactivate nucleases and consider bleach (sodium hypochlorite) for surface decontamination where appropriate.

Authoritative information: Always consult the product’s SDS and institutional EHS guidance.

Solvent Selection

Traditional organic solvent selection is not directly applicable to siRNA duplexes. Reconstitution and dilution should be performed in nuclease-free aqueous systems.

• Primary diluents (general, literature):
  • RNase-free water for initial dissolution.
  • TE or Tris buffer (e.g., 10 mM Tris-HCl, 0.1–1 mM EDTA, pH 7.5–8.0) to enhance stability during storage by chelating divalent cations.
  • For cell transfection, dilute into serum-free medium for complex formation with the chosen transfection reagent, then move to complete medium as recommended by the reagent supplier.
• Ionic strength and pH: Near-neutral pH (7.0–8.0) supports duplex integrity; avoid acidic conditions that promote hydrolysis.
• Avoid: Organic solvents (acetonitrile, DMSO) unless specifically validated for a particular delivery system; RNases; high temperatures.

Comparison (general):

• RNase-free water: maximal simplicity; best for short-term use and immediate transfection.
• TE buffer: improved long-term stability at -20 °C; EDTA sequesters Mg2+ that can catalyze RNA degradation.

Note: Exact solvent/buffer requirements for this item are not specified; refer to the CoA/Spec Sheet and your transfection reagent instructions.

Storage and Reconstitution

Storage (item-specific):

• Store at -20 °C.
• Shipped in: Ice chest + Ice pads.
• Research Use Note: For research use only.

Reconstitution and handling (general, literature; not item-specific specs):

• Initial reconstitution: Dissolve in RNase-free water or TE buffer to prepare a concentrated stock (e.g., 10–100 µM). Vortex gently and briefly spin down to collect solution.
• Aliquoting: Divide into single-use aliquots to avoid freeze–thaw cycles that can degrade RNA.
• Storage after reconstitution: -20 °C for medium-term (weeks–months) or -80 °C for long-term storage. TE buffer can enhance stability via EDTA chelation of Mg2+.
• Working solutions: Prepare immediately before use in nuclease-free water or serum-free medium for complex formation with the transfection reagent.
• Thawing: Thaw on ice; mix gently. Avoid extended time at room temperature and exposure to RNases.
• Concentration verification: If needed, quantify by A260 using RNase-free cuvettes and appropriate extinction coefficients (sequence-dependent; not provided for this item).

Any item-specific stabilizers, desalting/HPLC status, exact extinction coefficients, and recommended stock concentrations: Not specified for this item; refer to CoA/Spec Sheet.

Structure and Identity

Short interfering RNA (siRNA) duplexes targeting the human UQCRB transcript (ubiquinol–cytochrome c reductase binding protein, Complex III subunit). This product is a pre-designed set (“Set A”), typically comprising multiple non-overlapping siRNA duplexes directed against the same gene to increase the likelihood of robust knockdown across cell types.

Item-specific identifiers:

• Product Name: UQCRB Human Pre-designed siRNA Set A
• SKU: U1479554
• Target gene: UQCRB (Homo sapiens)
• CAS, CID, InChIKey, SMILES: Not applicable to short RNA duplex mixtures; not specified for this item; refer to CoA/Spec Sheet.

Structural features (general for siRNA; literature):

• Double-stranded RNA, usually 19–21 bp with 2-nt 3' overhangs (e.g., dTdT), sense/antisense strands with 5' phosphate/3' hydroxyl ends depending on synthesis and any stabilizers.
• Phosphodiester backbone; may include optional chemical modifications to enhance stability or reduce off-targeting (not specified for this item).
• No defined small-molecule stereocenters; chiral centers reside on ribose units along the polymeric backbone.

2D description in words (general):

• Two antiparallel RNA strands forming A-form helix segments; each nucleotide contains a ribose (typically 2'-OH) linked to heterocyclic bases (A, U, G, C) via N-glycosidic bonds; inter-nucleotide phosphodiester linkages connect 3'–5'.

Molecular formula and molecular weight:

• Not specified for this item; refer to CoA/Spec Sheet (values depend on exact sequences and any modifications).

Synthetic Utility

Not a small-molecule building block. As a functional genomics tool, this siRNA set has utility in synthetic biology and systems biochemistry experiments rather than in traditional chemical synthesis.

General research utility (literature):

• Target validation: Transiently suppress UQCRB to test pathway hypotheses, identify compensatory mechanisms, and prioritize targets for follow-up with CRISPR or small-molecule modulation.
• Chemical biology: Combine siRNA knockdown with small-molecule inhibitors to probe synthetic interactions, epistasis, or pathway redundancy (e.g., OXPHOS modulators).
• Omics integration: Pair RNAi perturbation with transcriptomics, proteomics, and metabolomics to map UQCRB-dependent networks.

Note: No item-specific synthetic or conjugation features (e.g., dyes, biotin, conjugates) are specified for this product.

Target Specificity

This section is primarily intended for antibodies; however, for clarity:

• Target: Human UQCRB mRNA (gene symbol provided in product name). Further identifiers (transcript variants, accession numbers, sequences) are not specified for this item.
• siRNA content: Pre-designed Set A typically includes multiple distinct duplexes for the same target; exact sequences, modifications, and design criteria are not specified for this item; refer to CoA/Spec Sheet.

No additional item-specific target validation data (e.g., tested cell lines, observed knockdown percentages) are provided.

Domande frequenti

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.