CUL3 Human Pre-designed siRNA Set A

Cat. No.: C1479705

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Vue d’ensemble

CUL3 Human Pre-designed siRNA Set A contains three designed siRNAs for CUL3 gene (Human), as well as a negative control, a positive control, and a FAM-labeled negative control. Components CUL3 siRNA-1: 5 nmol (HPLC) CUL3 siRNA-2: 5 nmol (HPLC) CUL3 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

Conditions de stockage de stockage

Store at -20°C

Expédié en

Ice chest + Ice pads

Ce produit nécessite l'expédition en chaîne froide. Les services terrestres et autres services économiques ne sont pas disponibles.

Product Line

Produits biologiques

Application Protocols

Item-specific, tested applications and protocols are not provided in the product data for SKU C1479705.

• Recommended action
  • Not specified for this item; refer to CoA/Spec Sheet.
  • Follow your transfection reagent manufacturer’s instructions and institutional SOPs for siRNA handling and transfection.

No validated dilutions, positive controls, or platform-specific protocols are supplied in the current listing.

Biological Roles

• Target overview: CUL3 (Cullin-3) encodes a core scaffold protein of CUL3–RBX1 Cullin–RING E3 ubiquitin ligase (CRL3) complexes.

• Mechanistic roles (literature)

  • Scaffold function: CUL3 bridges the RING protein RBX1 and BTB-domain substrate adaptors, positioning E2~Ub for substrate ubiquitylation.
  • Proteostasis: CRL3 complexes regulate proteasomal turnover of numerous substrates, influencing cell cycle, cytoskeletal dynamics, and stress responses.
  • Redox/homeostasis: Through BTB adaptor KEAP1, CRL3 targets NRF2 for ubiquitylation, modulating antioxidant response element (ARE)-driven transcription.
  • Signaling interplay: Impacts small GTPase regulation, ion channel trafficking, and mitotic control via adaptor-specific substrate selection.

• Research utility of CUL3 knockdown (general)

  • Enables transient suppression of CRL3 activity to dissect adaptor–substrate relationships.
  • Facilitates evaluation of NRF2 pathway dynamics, ubiquitination kinetics, and proteasomal dependency in diverse cell models.
  • Supports functional genomics screens examining dependencies on ubiquitin pathway components.

Note: The above roles summarize general biology of human CUL3. This product specifically targets CUL3 mRNA for research use; no medical or clinical claims are made.

Buffer Applications

Buffering here pertains to siRNA dissolution, storage, and transfection complex formation rather than classical analytical buffer systems.

• Reconstitution/storage buffers (general)

  • RNase-free water for initial dissolution.
  • TE or TE-low (10 mM Tris-HCl, pH 7.4–8.0; 0.1–1 mM EDTA) for longer-term stability by chelating trace metal ions.
  • Avoid Mg2+ and Ca2+ in storage buffers to limit RNA backbone cleavage.

• Working solutions (general)

  • Prepare concentrated stocks (e.g., 10–100 µM) and dilute into serum-free medium (e.g., Opti-MEM) for complex formation with lipid-based transfection reagents, followed by addition to complete medium.

• Practical tips

  • Use certified RNase/DNase-free consumables; clean work areas with RNase decontamination reagents.
  • Filter sterilize buffers if sterility is required; avoid autoclaving after adding EDTA or Tris.

Item-specific buffer recommendations are not provided; consult the datasheet or CoA for lot-specific guidance.

Green Alternatives

While “green chemistry” comparisons are less applicable to a nucleic acid reagent than to organic solvents or bulk chemicals, sustainability can still be improved by choices in buffers, consumables, and workflow design.

• Greener practice options (general)

  • Aqueous systems: Use RNase-free water and minimal buffer concentrations (e.g., TE-low) to reduce chemical footprint; avoid DEPC in favor of certified nuclease-free water to eliminate hazardous residues.
  • Minimize plastics: Consolidate aliquot sizes to minimize tube usage; opt for recycled-content tip boxes and recyclable packaging where validated for RNase-free work.
  • Energy efficiency: Store working aliquots at −20°C rather than −80°C when stability permits; consolidate freezers and maintain proper defrosting schedules.
  • Waste reduction: Plan transfection layouts to optimize well usage and reduce surplus complexes.

• Comparison (general)

  • DEPC-treated water vs certified nuclease-free water: The latter avoids handling carcinogenic DEPC and quenching steps, reducing hazards and time.
  • Ethanol-based cleanups vs spin-column salt exchange: Spin columns with aqueous buffers can reduce alcohol volumes, though assess RNA recovery and RNase risk.

These are general sustainability suggestions; no item-specific green credentials are provided in the product data.

Pharmaceutical Uses

This product is provided strictly for research use only and is not intended for diagnostic, therapeutic, or clinical applications.

• Formulation/excipient status

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

• General context (literature)

  • siRNA reagents for pharmaceutical applications require GMP manufacturing, stringent impurity controls (including truncated species, endotoxin, and residual solvents), and validated delivery systems; these aspects are outside the scope of this research-grade listing.

• Laboratory translation considerations

  • For preclinical research, maintain detailed records of lot numbers, purity, and analytical characterization to support reproducibility; however, no pharmacopeial status or clinical suitability is claimed or implied for this product.

Physical Properties

Item-specific physicochemical specifications are not provided for this siRNA set.

• Item-specific values

  • Appearance: Not specified for this item; refer to CoA/Spec Sheet.
  • Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
  • Extinction coefficient, OD260-to-mass conversion, residual salts/solvents: Not specified for this item; refer to CoA/Spec Sheet.

• General properties for siRNA oligonucleotides (literature)

  • Typical duplex length: 19–21 bp with 2-nt 3′ overhangs.
  • Approximate molecular mass: ~13–15 kDa for a 21-mer duplex (sequence-dependent).
  • Solubility: Freely soluble in RNase-free water and low-ionic-strength buffers (e.g., 10 mM Tris, pH 7.4–8.0); avoid divalent cations for long-term storage.
  • Charge: Polyanionic due to phosphate backbone; does not cross lipid membranes without a delivery reagent.
  • Stability: Stable when lyophilized at −20 to −80°C; in solution, stability improves at pH 7–8 with RNase-free conditions; repeated freeze–thaw should be avoided.
  • UV absorbance: Strong at 260 nm; concentration typically determined by A260 (1 A260 ≈ 40 µg/mL for single-stranded RNA; duplex-specific factors vary).

• Reconstitution concentration (general)

  • Common working stocks: 10–100 µM in RNase-free water or TE (10 mM Tris, 0.1–1 mM EDTA), prepared aseptically to minimize RNase contamination.

Quality and Grades

• Item-specific quality information

  • Grade/Purity: Not specified for this item; refer to CoA/Spec Sheet.
  • Any sequence, length, modification, or desalting/HPLC details are not provided in the product data and should be confirmed on the CoA/Spec Sheet.

• Understanding siRNA quality metrics (general, for context)

  • Purity: Typically reported by HPLC/UPLC; higher-purity preparations reduce off-targets and innate immune activation due to truncated species.
  • Identity: Confirmed by MS (MALDI-TOF/ESI-MS) and, when applicable, strand stoichiometry for duplexes.
  • Quantitation: Provided as nmol/OD260, mass, or A260; extinction coefficients may be sequence-specific.
  • Residual salts/solvents: Low levels of TEA or acetonitrile from RP-HPLC may be present in general practice; specific limits must come from the CoA.
  • Endotoxin/RNase: For cell-based work, low endotoxin and RNase-free status are desirable; verification is vendor- and lot-specific.

• Practical implications

  • For reproducible knockdown, consistent purity and accurate quantitation are crucial; mix-and-match between lots should be minimized without re-optimization.
  • If using sensitive primary cells, prefer high-purity, low-endotoxin siRNA. Confirm with item-specific documentation before critical experiments.

Reaction and Applications

As a pre-designed siRNA set for human CUL3, this product is intended for RNA interference experiments and related gene-silencing workflows in mammalian cells.

• Core application (general, literature)

  • Sequence-specific knockdown of CUL3 mRNA via RISC-mediated cleavage, enabling transient loss-of-function studies.

• Typical experimental uses

  • Pathway mapping: Probe roles of CUL3-containing Cullin–RING E3 ligase complexes in ubiquitin-mediated proteostasis.
  • Phenotypic assays: Assess cell cycle, oxidative stress responses, or substrate stability influenced by CUL3-regulated ubiquitylation.
  • Validation: Orthogonal confirmation of CRISPR/Cas9 results; rescue studies with siRNA-resistant cDNA.

• Practical guidance (general)

  • Start with 5–50 nM final siRNA; optimize per cell type and transfection reagent.
  • Assess knockdown at 24–72 h by qRT-PCR (mRNA) and 48–96 h by Western blot (protein), accounting for target protein half-life.
  • Include controls: non-targeting siRNA, positive knockdown control, and transfection reagent-only.
  • Pooling of multiple siRNAs targeting CUL3 can improve robustness and mitigate off-targets; verify cytotoxicity and innate immune activation markers when needed.

• Readouts and QC

  • Monitor on-target effects and rule out off-target phenotypes via dose–response, multiple independent siRNAs, and rescue experiments.

Note: The above are general guidelines; item-specific sequences, modifications, and recommended conditions are not provided and should be checked on the CoA/Spec Sheet.

Reaction Conditions

Classical organic reaction conditions are not applicable. For experimental planning, the relevant “conditions” pertain to transfection and knockdown workflows (general literature guidance; not item-specific):

• Dosing and timing

  • Start at 5–50 nM final siRNA in cell culture; optimize to minimize cytotoxicity while maximizing knockdown.
  • Assess mRNA reduction by qRT-PCR at 24–72 h; protein reduction typically at 48–96 h depending on target half-life.

• Complex formation

  • Use lipid-based transfection reagents per manufacturer instructions (e.g., mix in serum-free medium such as Opti-MEM, incubate 5–20 min to form complexes, then add to cells).
  • Maintain RNase-free technique; prepare fresh complexes for each experiment.

• Controls and validation

  • Include non-targeting siRNA control, positive control siRNA, and reagent-only control.
  • Validate on-target effects with at least two independent siRNAs or an siRNA pool; perform rescue with siRNA-resistant CUL3 cDNA where feasible.

• Readouts

  • Western blot for CUL3 or downstream substrates/adaptors; reporter assays for pathway activity (e.g., NRF2/ARE reporters) as appropriate.

These are general conditions for siRNA-mediated knockdown; item-specific recommendations, sequences, and modifications are not provided and should be obtained from the CoA/Spec Sheet.

Safety and Handling

• Regulatory and hazard status

  • Research Use: For research use only.
  • GHS classification, signal word, hazard statements, pictograms: Not specified for this item; refer to SDS.

• General safety for oligonucleotides (literature/guidance)

  • siRNA is non-volatile and generally considered of low acute toxicity; nonetheless, handle as a laboratory chemical.
  • Avoid inhalation of powders/aerosols and contact with eyes/skin; use appropriate PPE.

• Recommended PPE and engineering controls

  • Lab coat, nitrile gloves, and safety glasses.
  • Work in a clean area dedicated to RNA handling to minimize RNase contamination; use RNase-free consumables and filtered tips.

• Incompatibilities and stability considerations

  • Avoid RNase contamination (do not mouth pipette; clean benches/tools with RNase decontamination solutions).
  • Avoid repeated freeze–thaw cycles to prevent degradation; aliquot upon first reconstitution.
  • Strong acids/bases and nucleases degrade RNA.

• First-aid overview (general)

  • Skin/eye contact: Rinse with water for several minutes; remove contaminated clothing; seek medical attention if irritation persists.
  • Inhalation: Move to fresh air; seek medical attention if symptoms occur.
  • Ingestion: Rinse mouth; seek medical attention if unwell.

Always consult the product’s SDS and institutional EHS guidance for definitive safety information and local requirements.

Solvent Selection

This product is an RNA duplex reagent; conventional organic solvent selection is not typically applicable. Focus is on nuclease-free aqueous dissolution.

• Recommended media (general, literature)

  • Primary: RNase-free water for reconstitution and short-term stocks.
  • TE or TE-low (10 mM Tris-HCl pH 7.4–8.0, 0.1–1 mM EDTA) for improved stability during storage by chelating trace divalent cations.
  • Avoid Mg2+/Ca2+ in storage buffers; divalent cations can catalyze RNA cleavage.

• Delivery mixtures (cell culture, general)

  • For transfection, combine with lipid-based reagents in serum-free medium (e.g., Opti-MEM) during complex formation, then add to complete medium as recommended by the transfection reagent manufacturer.

• Notes

  • Do not dissolve or store siRNA in alcohols or strong organic solvents; maintain aqueous, RNase-free conditions.
  • Filter sterilization (0.22 µm) of buffers can be used if sterility is required; avoid heat sterilization.

When planning experiments, prioritize RNase-free water or TE buffers; see Storage & Reconstitution for item-specific storage guidance.

Storage and Reconstitution

• Item-specific storage and shipping

  • Storage Conditions: Store at −20°C.
  • Shipped In: Ice chest + ice pads.

• Item-specific reconstitution details

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

• General guidance (literature; not item-specific specifications)

  • Reconstitution: Dissolve lyophilized siRNA in RNase-free water to a convenient stock (e.g., 10–100 µM). Mix gently; avoid vigorous vortexing of small volumes.
  • Aliquoting: Prepare single-use aliquots to avoid freeze–thaw; label with concentration and date.
  • Storage of solutions: Store aliquots at −20°C (short-to-medium term) or −80°C (long term). Protect from repeated freeze–thaw.
  • Handling: Use RNase-free tubes and tips; keep solutions on ice during setup. If using TE, limit EDTA to ≤1 mM to avoid chelating essential cations during transfection (dilution typically mitigates this).

Always consult the product’s datasheet/CoA for any lot-specific instructions on reconstitution volume, buffer choice, and concentration.

Structure and Identity

Short description: This product is a pre-designed, duplex small interfering RNA (siRNA) reagent set targeting the human CUL3 transcript (Cullin-3). Sequence-level details are not provided in the product data.

• Item-specific identifiers

  • SKU: C1479705
  • Product name: CUL3 Human Pre-designed siRNA Set A
  • Target: Human CUL3 mRNA (gene symbol: CUL3)
  • Molecular formula: Not specified for this item; refer to CoA/Spec Sheet.
  • Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
  • CAS, CID, InChIKey, SMILES: Not applicable/Not specified for siRNA oligonucleotide mixtures.

• Structural features (general, literature)

  • siRNA typically consists of a 19–21 bp double-stranded RNA duplex with 2-nt 3′ overhangs, phosphodiester backbone, and ribose sugars (2′-OH).
  • Sense (passenger) and antisense (guide) strands are complementary; the antisense strand is loaded into RISC to direct sequence-specific mRNA cleavage.
  • 2D depiction: two antiparallel RNA strands with Watson–Crick base pairing (A:U, G:C), often with UU or dTdT 3′ overhangs (design-dependent).

• Notes on composition (general)

  • Pre-designed siRNA sets are commonly provided as multiple duplexes targeting distinct regions of the same transcript to increase the likelihood of robust knockdown while enabling off-target assessment. The exact number and sequences are not specified here.

• Identity confirmation practices (general)

  • Vendors typically confirm oligo mass by MALDI-TOF or ESI-MS and assess purity by HPLC/UPLC; functional identity is supported by in-cell knockdown performance. Refer to CoA/Spec Sheet for item-specific details.

Synthetic Utility

Not typically applicable. As a biological oligonucleotide reagent, this siRNA set is not used as a classical synthetic building block or reagent in organic synthesis.

• Relevant perspective (general)
  • In chemical biology workflows, siRNA serves as a functional tool for target validation and pathway dissection rather than as a reactant.
  • Any chemical modification of siRNA (e.g., stabilizing sugars/backbones) would be a manufacturing detail; no such item-specific modifications are provided in the product data.

Target Specificity

• Item-specific information (from product name)

  • Target gene: CUL3 (Cullin-3)
  • Species: Human
  • Reagent type: Pre-designed siRNA Set A (sequence details not provided)

• Notes

  • The siRNA set is intended to target human CUL3 mRNA for RNAi-mediated silencing. No additional details (e.g., individual siRNA sequences, target exon regions, or predicted off-target analysis) are provided in the product data. Refer to the CoA/Spec Sheet for any available sequence-level or design annotations.

Foire aux questions

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.