EIF3H Human Pre-designed siRNA Set A

Cat. No.: E1485473

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Visão geral

EIF3H Human Pre-designed siRNA Set A contains three designed siRNAs for EIF3H gene (Human), as well as a negative control, a positive control, and a FAM-labeled negative control. Components EIF3H siRNA-1: 5 nmol (HPLC) EIF3H siRNA-2: 5 nmol (HPLC) EIF3H 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

Condições de armazenamento de armazenamento

Store at -20°C

Enviado em

Ice chest + Ice pads

Este produto requer transporte de cadeia fria. Serviços terrestres e outros serviços econômicos não estão disponíveis.

Application Protocols

General protocol for transfecting cultured cells with EIF3H siRNA (optimize per cell line and transfection reagent; research use only):

• Preparation
  • Thaw an aliquot on ice. Briefly centrifuge to collect contents. Work RNase-free.
  • Reconstitute lyophilized siRNA to 20–50 µM in nuclease-free water or 1× TE. For solutions, confirm supplied concentration by A260 if needed.
• Complexation (example)
  • Dilute siRNA in serum-free medium to 2× the desired final concentration (e.g., 20–100 nM).
  • In a separate tube, dilute lipid/polymer transfection reagent per manufacturer’s instructions in the same medium.
  • Combine solutions gently; incubate 10–20 min at room temperature to form complexes.
• Transfection
  • Replace cell culture medium with complex mix to achieve 5–50 nM final siRNA. Cells should be 30–70% confluent.
  • Incubate 4–6 h, then replace with complete medium unless reagent supports continuous exposure.
• Post-transfection analysis
  • 24–48 h: Extract RNA, perform qRT–PCR for EIF3H and housekeeping genes.
  • 48–96 h: Assess protein levels by Western blot; evaluate phenotypes (e.g., polysome profiling) as appropriate.
• Controls
  • Non-targeting siRNA negative control; positive control siRNA (e.g., against a known essential gene) to verify transfection efficiency; mock control.
• Troubleshooting
  • Low knockdown: Titrate siRNA and reagent amounts; verify cell health; consider alternative transfection chemistries or electroporation.
  • Cytotoxicity: Reduce reagent amount or siRNA dose; shorten exposure time.

Biological Roles

• Target overview (literature/general)
  • EIF3H encodes subunit H of the eukaryotic translation initiation factor 3 (eIF3) complex, a multi-subunit assembly that promotes 40S ribosomal subunit recruitment to mRNA and facilitates pre-initiation complex formation in cap-dependent translation.
  • EIF3H contributes to stabilizing eIF3 architecture and interacts with other eIF3 subunits to regulate mRNA recruitment and scanning.
• Cellular processes
  • Translation initiation: EIF3H participates in assembly of the 43S pre-initiation complex and in mRNA triage on the 40S subunit (literature).
  • Proteostasis: By influencing translation efficiency, EIF3H levels can modulate the synthesis rates of numerous proteins.
• Experimental implications of knockdown (general research context)
  • Reducing EIF3H mRNA via siRNA typically leads to diminished EIF3 complex function and can decrease global cap-dependent translation; the magnitude depends on cell type, expression level, and compensatory mechanisms.
  • Phenotypic readouts may include altered polysome profiles, slowed cell-cycle progression, or stress response activation markers in some systems (research observations; verify empirically).
• Orthology/annotations (reference only)
  • Species: Homo sapiens target. Rodent orthologs exist but sequence divergence requires species-specific siRNA designs (literature).

Buffer Applications

This product is not a buffering reagent. However, appropriate buffers are essential for siRNA stability and use.

• Recommended stock buffer systems (general)
  • 1× TE, pH 7.5–8.0 (10 mM Tris-HCl, 1 mM EDTA): Common for long-term storage stocks; EDTA chelates divalent cations that can promote RNA hydrolysis.
  • Nuclease-free water: Suitable for immediate use and short-term storage; minimize freeze–thaw by aliquoting.
• Handling tips
  • Avoid phosphate buffers with high Mg2+/Ca2+ during stock preparation; divalent cations increase cleavage rates at elevated temperatures.
  • Filter sterilization is typically unnecessary if using certified nuclease-free components; avoid exposing siRNA to shear (vortex gently or flick; brief low-speed spin-down).
• Working solutions
  • For transfection, dilute stocks into serum-free transfection medium according to the transfection reagent protocol, then replace with complete medium after complexation/incubation as directed.

Green Alternatives

Although this is a biological reagent rather than a solvent/chemical process intermediate, greener practices can be applied to its use.

• Transfection and workflow choices (general)
  • Prefer biodegradable, low-toxicity transfection reagents to reduce hazardous waste and improve cell viability (evaluate performance empirically).
  • Minimize single-use plastics by planning aliquots and consolidating setups; choose low-retention, recyclable plastics where permitted by RNase-free requirements.
  • Use room-temperature stable ancillary reagents when feasible to reduce cold-chain energy demands (without compromising siRNA integrity, which still requires −20°C storage as specified).
• Process optimization to reduce waste
  • Design experiments with factorial/DoE approaches to minimize screening runs while identifying optimal siRNA dose and reagent ratios.
  • Pool analysis: Start with pooled siRNA to determine phenotype, then deconvolute with individual duplexes only if needed.
• Comparative considerations
  • CRISPR interference (CRISPRi) can be a non-templated, renewable alternative for gene suppression in some models, reducing repeated siRNA consumption; however, it requires genetic engineering and may alter chromatin state. siRNA remains advantageous for rapid, transient knockdown without genomic modification (literature).

Pharmaceutical Uses

This product is supplied strictly for research use only (per product data) and is not intended for human or animal therapeutic, diagnostic, or clinical applications.

• Research/manufacturing context (general)
  • In discovery biology, siRNA reagents are widely used for target validation, pathway dissection, and mechanism-of-action studies to support early-stage screening campaigns.
  • In bioprocess development, transient knockdown can be used to probe host-cell protein expression impacts on product quality attributes (experimental context only).
• Pharmacopeial/excipient status
  • No pharmacopeial monograph or excipient designation is applicable to this siRNA research reagent.
• Compliance notes
  • Users are responsible for ensuring institutional approvals for work with recombinant nucleic acids and for adhering to local regulations governing disposal of nucleic acid materials and transfection reagents.

Physical Properties

• Item-specific properties
  • Appearance: Not specified for this item; refer to CoA/Spec Sheet.
  • Concentration/formulation: Not specified for this item; refer to CoA/Spec Sheet.
  • Solubility: Readily soluble in nuclease-free water or 1× TE buffer (typical for siRNA; item-specific solubility not specified; refer to CoA/Spec Sheet).
  • pH of supplied solution: Not specified for this item; refer to CoA/Spec Sheet.
• Literature/general properties for siRNA (not item-specific)
  • Physical state: Dry (lyophilized) or aqueous solution; hygroscopic when dry.
  • Solubility: Highly soluble in aqueous buffers; insoluble in nonpolar organic solvents.
  • UV absorbance: Peak near 260 nm; typical A260 of 1.0 corresponds to ~40 µg/mL single-stranded RNA; duplex extinction coefficients depend on sequence/length (literature).
  • Thermal stability: Duplex melting temperatures (Tm) commonly 60–85°C depending on sequence/length/ionic strength (literature). Salts (Na+, K+, Mg2+) raise Tm.
  • LogP, refractive index, BP/MP: Not applicable in conventional sense for polyanionic biopolymers; not determined.
• Practical notes
  • Reconstitution in RNase-free water or 1× TE (10 mM Tris-HCl, 1 mM EDTA, pH 7.5–8.0) is standard. EDTA chelates trace divalent cations that can promote RNA cleavage (literature).
  • Avoid repeated freeze–thaw; duplex integrity can be monitored by native PAGE or HPLC (general guidance).

Quality and Grades

• Item-specific quality declarations
  • Grade/purity: Not specified for this item; refer to CoA/Spec Sheet.
  • Stabilizers/modifications: Not specified for this item; refer to CoA/Spec Sheet.
• What “pre-designed siRNA set” typically means (literature/general)
  • Multiple distinct duplexes targeting non-overlapping regions of the EIF3H transcript, intended to be used individually or as a pooled mix to enhance knockdown efficiency and mitigate sequence-specific off-targets.
  • Typical QC for research-grade siRNA includes OD260 quantitation, desalting or HPLC/PAGE purification, and identity confirmation by MALDI-TOF or LC-MS (varies by supplier). These details are not specified for this item; refer to CoA/Spec Sheet.
  • Low endotoxin and RNase-free manufacturing are commonly expected for cell culture use; specific limits are not specified for this item; refer to CoA/Spec Sheet.
• Practical implications
  • If using for sensitive phenotypic assays, HPLC- or PAGE-purified siRNA can reduce truncated species (general). Verify purification grade on the CoA.
  • For comparative studies across targets, normalize by nmol amount and confirm actual concentration by A260.
  • If the set is supplied lyophilized, note the stated nmol per tube on the label/CoA to calculate reconstitution volumes accurately.

Reaction and Applications

• Primary application (item-specific context)
  • Pre-designed siRNA set targeting human EIF3H for transient knockdown in cultured cells. Intended for research use only.
• Mechanistic background (literature)
  • siRNA duplexes are incorporated into RISC, the passenger strand is discarded, and the guide strand directs Argonaute-catalyzed cleavage or translational repression of complementary EIF3H mRNA, reducing protein levels.
• Use cases (general)
  • Target validation: Confirm EIF3H involvement in translation initiation-dependent phenotypes.
  • Pathway mapping: Dissect eIF3 complex contributions to cap-dependent initiation and 40S recruitment.
  • Synthetic lethality screens: Combine EIF3H knockdown with genetic/chemical perturbations (research context).
• Practical tips
  • Test duplexes individually and as a pool; pooling can increase on-target knockdown while diluting sequence-specific off-targets.
  • Optimize transfection conditions for each cell line: reagent type, siRNA dose (typical 5–50 nM), cell confluency (30–70%), and exposure time.
  • Verify knockdown by qRT–PCR at 24–48 h and by immunoblot at 48–96 h (depending on EIF3H protein half-life; literature).
  • Include controls: non-targeting siRNA, positive control siRNA (e.g., against PLK1) to benchmark transfection efficiency, and mock controls.
  • For longer studies, consider sequential dosing (e.g., day 0 and day 3) to sustain suppression in dividing cells (general guidance).

Reaction Conditions

Classical chemical reaction conditions are not applicable. For experimental conditions relevant to siRNA-mediated knockdown, see below (general guidance; optimize per cell model and transfection reagent):

• Complex formation (literature/general)
  • Mix siRNA (final 5–50 nM) with the recommended volume of lipid/polymer transfection reagent in serum-free medium (e.g., Opti-MEM). Incubate 10–20 min at room temperature to allow complexation.
• Cell conditions
  • Seed cells to achieve ~30–70% confluency at transfection time. Replace culture medium with complex-containing mixture and incubate per reagent protocol (commonly 4–6 h) before switching to complete medium.
• Incubation and readouts
  • mRNA assessment by qRT–PCR: 24–48 h post-transfection.
  • Protein level assessment by Western blot/ELISA: 48–96 h, depending on protein turnover.
• Sequential treatment
  • For sustained knockdown in rapidly dividing cells, a second dose at 48–72 h may improve and extend suppression.
• Controls and benchmarks
  • Include non-targeting negative control, positive control siRNA, and transfection reagent–only control to interpret on-/off-target effects and cytotoxicity.

Safety and Handling

• Regulatory and hazard data
  • GHS classification: Not specified for this item; refer to SDS.
  • Signal word: Not specified for this item; refer to SDS.
  • H-statements/pictograms: Not specified for this item; refer to SDS.
• General safety (literature/best practice for siRNA)
  • siRNA duplexes are generally considered low hazard for laboratory handling; treat as laboratory chemical/biological material. Avoid inhalation of powders and aerosol generation.
  • Use appropriate PPE: lab coat, gloves (change frequently to avoid RNase contamination), and safety glasses.
  • Work in a clean, RNase-free area. Use RNase-free consumables; wipe benches and pipettes with RNase decontamination solution.
  • Incompatibilities: Strong acids/bases and RNases; avoid divalent metal contamination (e.g., Mg2+, Mn2+) that may catalyze RNA degradation at elevated temperatures.
• First aid (overview; defer to SDS)
  • Skin/eye contact: Rinse with water. Remove contaminated clothing. Seek medical attention if irritation persists.
  • Ingestion/inhalation: Move to fresh air, rinse mouth; seek medical advice as appropriate.
• Spills/waste
  • Collect with RNase-free wipes; dispose per institutional nucleic acid waste procedures.
• Transport/storage precautions
  • Shipped: Ice chest + ice pads (per product data).
  • Storage: −20°C (per product data). Protect from moisture and RNases. Store in low-adsorption tubes. Avoid repeated freeze–thaw by aliquoting upon first thaw.

Solvent Selection

This product is an RNA duplex reagent and is not used as an organic solvent; instead, appropriate aqueous media are chosen for reconstitution and delivery.

• Recommended reconstitution media (general/literature)
  • Nuclease-free water (RNase/DNase-free): Minimizes ionic interactions; suitable for short-term storage and immediate transfection.
  • 1× TE buffer (10 mM Tris-HCl, 1 mM EDTA, pH 7.5–8.0): EDTA chelates divalent cations to limit hydrolysis during storage; commonly used for long-term stock solutions.
• Working solution/delivery media
  • Prepare working dilutions in serum-free, antibiotic-free transfection medium (e.g., Opti-MEM or equivalent) for complexation with transfection reagents (literature). After 4–6 h, replace with complete growth medium as per transfection reagent guidance.
• Compatibility notes
  • Avoid strong acids/bases and organic solvents; siRNA is insoluble/unstable in most nonaqueous media.
  • Presence of Mg2+ can promote backbone cleavage at elevated temperatures; keep Mg2+ low during stock preparation unless complexing with cationic lipids/polymers.
• Practical comparison (general)
  • Water vs TE: Water is ideal for immediate use; TE offers improved stability for storage but EDTA may interfere with cation-dependent transfection systems if not sufficiently diluted. Ensure ≥1:20 dilution of TE in final transfection mixes.

Storage and Reconstitution

• Storage conditions (item-specific)
  • Store at −20°C (per product data). Protect from moisture and RNases. Shipments are provided in an ice chest with ice pads.
• Reconstitution (general guidance for siRNA; verify with CoA/label)
  • Lyophilized material: Spin briefly, then add nuclease-free water or 1× TE to achieve a convenient stock concentration (e.g., 20–50 µM). Mix gently; avoid vigorous vortexing; allow 10 min at room temperature for full dissolution.
  • Solutions (if supplied): Upon receipt, aliquot into low-adsorption, RNase-free tubes to minimize freeze–thaw.
• Aliquoting and handling
  • Prepare single-use aliquots sized for one experiment to prevent repeated freeze–thaw cycles.
  • Keep on ice during setup; return unused aliquots promptly to −20°C.
• Stability (literature/general)
  • At −20°C, siRNA stocks in 1× TE are typically stable for ≥6 months; at −80°C, longer-term storage is common. Working dilutions at 4°C should be used within 1–2 weeks. Actual guaranteed stability for this item: Not specified for this item; refer to CoA/Spec Sheet.
• Thawing/refreezing
  • Avoid multiple freeze–thaw cycles; if unavoidable, limit to ≤3 and assess integrity by gel or analytical HPLC if critical.
• RNase control
  • Use certified RNase-free tips/tubes and dedicated solutions. Clean work surfaces with RNase decontamination reagents before handling.

Structure and Identity

This product is a pre-designed small interfering RNA (siRNA) set targeting human EIF3H (Eukaryotic translation initiation factor 3 subunit H) for gene knockdown applications.

• Item-specific identifiers
  • SKU: E1485473
  • Product name: EIF3H Human Pre-designed siRNA Set A
  • Category: 小分子和化合物库 (research reagent; siRNA pool)
• Item-specific structural data
  • 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: Not applicable for oligonucleotide duplexes; Not specified for this item; refer to CoA/Spec Sheet.
  • InChIKey: Not applicable for oligonucleotide duplexes; Not specified for this item; refer to CoA/Spec Sheet.
• Literature/general description of structure (not item-specific)
  • siRNA typically consists of 19–21 bp RNA duplexes with 2-nt 3′ overhangs on each strand, bearing 5′-phosphate/3′-hydroxyl termini (literature).
  • Chemical modifications (e.g., 2′-OMe, phosphorothioate) may be employed in some siRNAs to enhance stability; presence/absence for this item: Not specified for this item; refer to CoA/Spec Sheet.
  • Target: Human EIF3H mRNA; pre-designed sets usually comprise multiple non-overlapping duplexes to maximize knockdown robustness (literature).

Synthetic Utility

Not typically applicable. This is a biological gene-silencing reagent (siRNA) rather than a small-molecule building block or catalyst. It does not serve as a reagent for organic synthesis.

• For users seeking chemical synthesis tools
  • Consider small-molecule translation inhibitors or ribosome-targeted probes if the experimental goal is chemical perturbation of translation initiation rather than genetic knockdown.
  • For bioconjugation workflows, siRNA can be chemically modified (e.g., 5′-thiol, amine, azide) for coupling to carriers; however, modification status for this item is Not specified for this item; refer to CoA/Spec Sheet.

Target Specificity

• Item-specific target
  • Gene: EIF3H (Eukaryotic translation initiation factor 3 subunit H), Homo sapiens.
  • Intended action: Sequence-specific RNAi-mediated suppression of EIF3H mRNA.
  • Duplex content/sequences: Not specified for this item; refer to CoA/Spec Sheet.
• General notes on specificity (literature)
  • Pre-designed siRNA sets typically include multiple duplexes targeting distinct regions of the transcript to mitigate off-target effects and sequence-dependent variability.
  • Off-targets can arise from seed-region partial complementarity; using a pooled set and validating phenotype with multiple independent duplexes helps confirm on-target activity.
  • Use strand-specific qPCR or 5′-RACE assays, if needed, to verify on-target cleavage products.