CSK Human Pre-designed siRNA Set A

Cat. No.: C1479552

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Visão geral

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

Product Line

Produtos biológicos

Application Protocols

Tested/validated application details are not specified for this item; refer to product datasheet/CoA. The following is general guidance for siRNA transfection (literature/general):

• Rapid protocol (adherent cells, 24-well):
  1. Plate 5–8 × 10^4 cells/well to reach ~60% confluence next day.
  2. Dilute siRNA to desired working concentration (e.g., 5–20 nM final) in 50 µL serum-free medium.
  3. Dilute transfection reagent separately in 50 µL serum-free medium (amount per supplier’s guidance).
  4. Combine, mix gently, and incubate 10–20 min at room temperature.
  5. Add complexes dropwise to cells in 400 µL medium (serum-free or reduced-serum as recommended).
  6. After 4–6 h, replace with 500–1000 µL complete medium if required.
  7. Assess mRNA at 24–48 h; protein at 48–96 h.
• Controls and verification:
  • Include non-targeting and positive-control siRNAs; verify CSK knockdown by RT-qPCR and Western blot.
• Notes:
  • Optimize dose and reagent for each cell line; monitor cytotoxicity and adjust conditions accordingly.

Biological Roles

• Target gene overview (literature/general):
  • CSK (C-terminal Src kinase) is a cytoplasmic tyrosine kinase that phosphorylates the conserved C-terminal inhibitory tyrosine of Src family kinases (SFKs), switching them to a closed, inactive conformation.
  • Through SFK suppression, CSK influences focal adhesion dynamics, actin cytoskeleton organization, integrin signaling, GPCR and growth factor signaling, and downstream MAPK/ERK pathways.
• Cellular context:
  • CSK lacks a membrane-targeting myristoylation motif and relies on adaptors (e.g., Cbp/PAG) for localization to membrane microdomains where SFKs operate.
  • CSK activity maintains signaling homeostasis; reduced CSK function generally leads to hyperactive SFKs and enhanced tyrosine phosphorylation cascades.
• Experimental implications for RNAi knockdown (literature/general):
  • Depletion of CSK typically increases SFK activity markers (e.g., pY416-Src), alters adhesion turnover, and can modulate proliferation, migration, and survival phenotypes in cell-type–specific ways.
  • Combines synergistically with pharmacologic SFK inhibitors to probe pathway wiring and feedback regulation.
• Systems perspective:
  • CSK is a key node in network robustness, providing negative feedback to multiple receptor inputs, making it a valuable target for pathway dissection in diverse human cell models.

Buffer Applications

This product is not a chemical buffering agent; however, appropriate buffers are essential for handling and storing siRNA.

• Stock solution preparation (literature/general):
  • RNase-free water (preferred for maximal flexibility) or TE buffer, pH 7.5–8.0 (10 mM Tris-HCl, 0.1–1 mM EDTA) for enhanced stability by chelating Mg2+ that supports nuclease activity.
  • Typical stock concentration: 10–100 µM; select based on anticipated working range to minimize dilutions.
• Working solutions for delivery:
  • Prepare in serum-free, RNase-free medium recommended for the transfection reagent (e.g., Opti-MEM), then add to cells; restore serum after complexation per reagent protocol.
• Practical notes:
  • Use certified RNase-/DNase-free consumables.
  • Filter-sterilize buffers if prepared in-house; avoid autoclaving solutions containing EDTA with certain plastics that may leach.
  • Avoid phosphate buffers during some cationic polymer transfections where phosphate can interfere with complex formation.

Green Alternatives

While siRNA reagents are aqueous and generally low in environmental hazard, greener practices can reduce waste and hazardous inputs in RNAi workflows.

• Greener choices (literature/general):
  • Use RNase-free water in place of DEPC-treated water to avoid handling toxic DEPC, relying on certified nuclease-free consumables.
  • Prefer low-toxicity, serum-compatible transfection reagents that allow delivery in reduced volumes and at lower siRNA doses, decreasing reagent consumption and cell culture waste.
  • Employ miniaturized, high-throughput formats (96–384 well) to reduce plastic and reagent usage.
• Waste reduction:
  • Aliquot concentrated stocks to extend shelf life and avoid repeated freeze–thaw and discard cycles.
  • Use refillable reservoirs and multi-channel pipetting to minimize tips and reservoirs per experiment.
• Energy considerations:
  • Consolidate freezer space and use −70 to −80 °C storage for long-term aqueous stocks only when necessary; retain lyophilized materials at −20 °C as feasible per stability guidance.
• Trade-offs:
  • Serum-free complexation steps reduce protein waste but may transiently stress cells; brief serum starvation windows should be minimized to maintain viability.

Pharmaceutical Uses

This product is supplied strictly for research use only. No medical, diagnostic, or therapeutic use is intended or permitted.

• Research/formulation context (literature/general):
  • Preclinical tool for evaluating delivery technologies (lipids, polymers, peptides) and assessing endosomal escape and intracellular trafficking using a defined gene target (CSK).
  • Benchmarking siRNA pharmacology in cell culture for dose–response, duration of knockdown, and off-target assessment.
• Regulatory notes:
  • siRNA products intended for clinical or GMP manufacturing must adhere to pharmacopeial and GMP standards (identity, purity, residual solvents, endotoxin, sterility). Those requirements are beyond the scope of this research-grade product.

Physical Properties

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight (per duplex): Not specified for this item; depends on sequence length and composition.
• Molecular formula: Not specified for this item.
• Solubility (literature/general for siRNA):
  • Freely soluble in RNase-free water or TE buffer (typically to 10–100 µM stock solutions).
  • Insoluble in nonpolar organic solvents; aqueous only.
• pKa / logP / refractive index: Not applicable for polyanionic nucleic acids in aqueous systems (literature/general).
• UV properties (literature/general):
  • Strong absorption at 260 nm; use A260 for quantification. Typical extinction coefficients are sequence-dependent.
• Physical state on receipt:
  • Often supplied lyophilized or in aqueous buffer; Not specified for this item.
• Stability (literature/general):
  • Stable for months at −20 °C when lyophilized; aqueous solutions benefit from aliquoting and storage at −80 to −20 °C to avoid freeze–thaw.
• Density / BP / MP: Not applicable/not meaningful for oligomer mixtures; not specified for this item.

Quality and Grades

• Item-specific grade/purity: Not specified for this item; refer to CoA/Spec Sheet.
• What “pre-designed siRNA set” typically implies (literature/general):
  • Multiple non-overlapping siRNA duplexes targeting the same gene (CSK) to increase likelihood of robust knockdown and enable pooled or individual testing.
  • Sequences are computationally selected to reduce off-targets and immunostimulation; exact criteria vary by supplier.
• Common purification levels for siRNA (literature/general):
  • Desalted (crude) vs. HPLC- or PAGE-purified. HPLC/PAGE improves length homogeneity and reduces n-1 species, often yielding more consistent knockdown and lower off-target effects.
• Stabilizers and formulation:
  • Some siRNAs are delivered lyophilized with volatile buffers or sugars (e.g., trehalose) to aid resuspension and stability; Not specified for this item.
• Quality control metrics you may expect on a CoA (for guidance only):
  • Assignment/purity by HPLC or CE, mass confirmation (MALDI-TOF/ESI), annealing verification, endotoxin level for cell-based assays, residual solvents/salts, and concentration after reconstitution.
• Recommendation:
  • Consult the product’s CoA/Spec Sheet for purification method, sequence information (if disclosed), and QC criteria specific to SKU C1479552.

Reaction and Applications

This siRNA set is designed for RNA interference (RNAi)–mediated gene silencing of human CSK.

• Core application (item intent):
  • Transient knockdown of CSK mRNA to study Src family kinase regulation, signaling pathways, and phenotypic outcomes in human cells.
• Experimental use-cases (literature/general):
  • Pathway analysis: Dissect CSK’s role in negative regulation of SFKs and downstream MAPK/FAK signaling.
  • Phenotypic screens: Use pooled siRNAs for robust primary screening; deconvolute with individual duplexes.
  • Target validation: Combine with pharmacological inhibitors/CRISPR to corroborate CSK dependency.
  • Rescue experiments: Co-transfect siRNA with siRNA-resistant CSK cDNA to confirm on-target effects.
• Practical considerations:
  • Use multiple siRNAs and include non-targeting and positive-control siRNAs (e.g., PLK1) to benchmark transfection and knockdown.
  • Quantify knockdown by RT-qPCR (mRNA) at 24–48 h and by immunoblotting (protein) at 48–96 h.
  • Titrate siRNA (e.g., 1–50 nM final, cell-type dependent) and optimize lipid/polymer ratios for maximal efficacy with minimal cytotoxicity.
  • Minimize off-targets by avoiding excessive concentrations and by using chemically modified siRNAs if available (not specified for this item).

Reaction Conditions

Chemical reaction conditions are not applicable; below are general experimental conditions for siRNA-mediated transfection and knockdown (literature/general):

• Cell plating:
  • Plate cells to reach 40–70% confluence at time of transfection; optimize density for each line to balance proliferation and uptake.
• siRNA dosing:
  • Typical final concentration range: 1–50 nM. Start with 5–10 nM for many adherent lines; titrate downward to minimize off-targets while retaining efficacy.
• Complex formation:
  • Dilute siRNA and transfection reagent separately in serum-free medium (e.g., Opti-MEM), combine, incubate 5–20 min at room temperature to form complexes.
  • Typical reagent ratios: per vendor instructions; optimize nitrogen/phosphate (N/P) for polymers or µL reagent per pmol siRNA for lipids.
• Incubation and media:
  • Add complexes to cells in reduced-serum or serum-free conditions for 2–6 h, then replace with complete medium unless reagent is serum-compatible.
• Timeline for readouts:
  • mRNA knockdown: assess 24–48 h by RT-qPCR.
  • Protein knockdown: assess 48–96 h by immunoblot or flow cytometry; extend to 5–7 days for long-lived proteins.
• Controls:
  • Include non-targeting siRNA, mock transfection, and positive-control siRNA; test at identical concentrations.
• Replicates and statistics:
  • Use biological triplicates and appropriate normalization (housekeeping genes for qPCR; loading controls for WB).

Safety and Handling

• GHS classification, signal word, pictograms, H-statements: Not specified for this item; refer to SDS.
• General hazard profile (literature/general for siRNA):
  • Non-volatile, non-flammable aqueous oligonucleotide; typically low acute toxicity. Handle as a laboratory chemical with standard precautions.
• PPE and hygiene:
  • Wear lab coat, gloves, safety glasses. Work in a clean, RNase-free area to prevent degradation. Change gloves frequently to avoid RNase contamination.
• Incompatibilities and handling notes:
  • Avoid RNase contamination (do not mouth-pipette; use RNase-free plastics and solutions). DEPC-treated water should be fully hydrolyzed before use if applicable.
  • Avoid strong acids/bases and nucleases which degrade RNA.
• First aid (general):
  • Skin/eye contact: Rinse with water. Inhalation of powders/aerosols: move to fresh air. Ingestion: rinse mouth. Seek medical attention per institutional policy.
• Environmental precautions:
  • Dispose of aqueous nucleic acid waste according to institutional biosafety and chemical waste guidelines. Avoid release to drains if local rules restrict nucleic acid disposal.
• Shipping and storage conditions (item-specific):
  • Shipped: Ice chest + ice pads (provided).
  • Storage: Store at −20 °C (per Product Data). Defer to SDS/CoA for any additional constraints.

Solvent Selection

For siRNA, “solvent” considerations mean aqueous buffers for reconstitution and working solutions rather than organic solvents.

• Recommended vehicles (literature/general):
  • RNase-free water for initial dissolution; TE buffer (10 mM Tris-HCl, 0.1–1 mM EDTA, pH 7.5–8.0) for enhanced stability in stock solutions.
  • For transfection complexation, dilute in serum-free, RNase-free cell culture medium (e.g., Opti-MEM) per transfection reagent guidance.
• Avoid:
  • Nonaqueous organic solvents; strong acids/bases; buffers containing RNases; repeated freeze–thaw in aqueous solution.
• Practical tips:
  • Warm to room temperature before opening to minimize condensation. Vortex gently or pipette to aid dissolution; do not heat above ambient unless supplier allows.
• Comparison (general):
  • RNase-free water: maximal flexibility for downstream buffers; slightly less long-term stability than TE.
  • TE buffer: improved stability via chelation of divalent cations; EDTA may interfere with divalent cation–dependent transfection chemistries. Choose based on your delivery method.

Storage and Reconstitution

• Item-specific storage: Store at −20 °C (per Product Data). Shipped in ice chest with ice pads.
• Reconstitution: Not specified for this item; refer to CoA/Spec Sheet. General guidance (literature):
  • Equilibrate vial to room temperature before opening to avoid condensation.
  • Reconstitute in RNase-free water or TE buffer to 10–100 µM. Mix gently until fully dissolved.
  • Spin down briefly to collect contents; avoid vigorous vortexing if foaming occurs.
• Aliquoting and stability (literature/general):
  • Prepare single-use aliquots to prevent repeated freeze–thaw.
  • Storage recommendations: Lyophilized at −20 °C (months). In solution: −80 to −20 °C for long-term (weeks–months depending on buffer), 4 °C for short-term (≤1 week). Avoid >3 freeze–thaw cycles.
  • Protect from RNases; use nuclease-free tubes and tips.
• Thawing and handling:
  • Thaw on ice or at room temperature; keep on ice during setup. Return unused aliquots to freezer promptly.
• Documentation:
  • Record reconstitution buffer, concentration, and aliquot map. Consult the product’s CoA/Spec Sheet for any item-specific stabilizers or recommended diluents.

Structure and Identity

This product is a gene-specific small interfering RNA (siRNA) set designed for human CSK (C-terminal Src kinase) knockdown. It comprises synthetic, double-stranded RNA oligonucleotides.

• Item-specific identifiers:
  • 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 / InChIKey / CAS / CID: Not applicable for RNA mixtures; not specified for this item.
• Composition and architecture (literature/general for siRNA):
  • Typical structure: 19–21 bp duplex with 2-nt 3′ overhangs (commonly dTdT or rUrU).
  • Strands: sense (passenger) and antisense (guide) RNA, annealed.
  • Chemical modifications: May be unmodified or contain stability-enhancing modifications (e.g., 2′-OMe, 2′-F, phosphorothioate) depending on design; Not specified for this item.
• Target identity (item intent):
  • Gene: CSK (C-terminal Src kinase), Homo sapiens.
  • Intended purpose: Post-transcriptional silencing of CSK mRNA via RISC-mediated cleavage.
• 2D description (general):
  • Linear polyanionic biopolymer strands composed of ribonucleotides (A, U, G, C) with 3′–5′ phosphodiester linkages forming a Watson–Crick base-paired duplex.

Synthetic Utility

This product is a biological knockdown reagent and does not serve as a synthetic building block or catalyst in organic/medicinal chemistry.

• Not typically applicable:
  • No role in stoichiometric or catalytic chemical synthesis, cross-coupling, or small-molecule transformations.
• Relevant redirection:
  • For practical use, see Reaction & Applications, Reaction Conditions, and Application Protocols (general guidance) for gene-silencing experiments.

Target Specificity

• Item-specific details available: Target gene only.
  • Target: CSK (C-terminal Src kinase), human.
  • Species reactivity: Human (intended target). Cross-reactivity to other species is sequence-dependent and not specified for this item.
  • siRNA sequences, modifications, and off-target assessments: Not specified for this item; refer to CoA/Spec Sheet if available.
• General notes (literature/general):
  • Pre-designed siRNA sets typically include multiple independent duplexes to mitigate single-sequence off-target effects and to enable pooled use for robust knockdown.
  • Off-target minimization strategies include seed-region deconvolution and concentration titration; apply validation via rescue experiments.

Perguntas frequentes

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.