Stat6 Rat Pre-designed siRNA Set A

Cat. No.: S1479081

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Panoramica

Stat6 Rat Pre-designed siRNA Set A contains three designed siRNAs for Stat6 gene (Rat), as well as a negative control, a positive control, and a FAM-labeled negative control. Components Stat6 siRNA-1: 5 nmol (HPLC) Stat6 siRNA-2: 5 nmol (HPLC) Stat6 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.

Application Protocols

Item-specific, tested applications: Not specified for this item. The following is general, literature-based guidance for transfecting siRNA into rat cells (not item-specific):

A. Lipid-mediated transfection (adherent cell line)

• Prepare stocks: Reconstitute siRNA in RNase‑free water (e.g., 20–100 µM). Dilute to working solutions on ice.
• Complexing: In separate tubes, dilute siRNA and transfection reagent in serum‑free medium (e.g., Opti‑MEM). Combine to achieve 5–50 nM final siRNA; incubate 10–20 min at RT.
• Cells: Plate cells to 40–60% confluence. Replace medium with fresh complete medium if reagent is serum‑compatible; otherwise add complexes in serum‑free medium and switch to complete medium after 4–6 h.
• Incubate 24–72 h. Assess mRNA (24–48 h) and protein (48–96 h) knockdown.

B. Electroporation (primary cells)

• Resuspend cells in electroporation buffer at vendor‑recommended density.
• Mix with siRNA to 100–300 nM in cuvette; pulse with preset program optimized for the cell type. Recover in prewarmed complete medium.

Controls and QC

• Include non‑targeting siRNA, a positive knockdown control, and a fluorescent uptake control if available.
• Verify knockdown by qPCR and Western blot; use STAT‑responsive reporter assays as functional confirmation.

Note: Optimize conditions for each cell type and transfection reagent. Item‑specific instructions are Not specified for this item.

Biological Roles

Target overview (general biology; not item-specific):

• Stat6 encodes Signal transducer and activator of transcription 6, a latent cytoplasmic transcription factor activated downstream of IL‑4 and IL‑13 receptors via JAK kinases.
• Upon cytokine stimulation, STAT6 is tyrosine‑phosphorylated, dimerizes, translocates to the nucleus, and binds GAS‑like DNA elements to regulate transcription.
• Key processes: differentiation of Th2‑like responses, alternative activation programs in macrophages, regulation of IgE class switch machinery, and induction of mucus‑associated genes in epithelial cells (rodent/human literature).
• Crosstalk: interfaces with STAT3/STAT5 pathways and PI3K/AKT signaling; cooperative and antagonistic promoter occupancy depends on cellular context.

Why Stat6 siRNA is used (research rationale; not item-specific):

• To acutely suppress Stat6 expression for temporally controlled pathway dissection without stable genome alteration.
• To verify on‑target effects seen with pharmacologic modulators of the IL‑4Rα/JAK pathway.
• To map STAT6‑dependent transcriptomes via RNA‑seq/ATAC‑seq and to assess changes in chromatin accessibility at STAT6 motifs.

Species note: This product targets rat Stat6 per item name; sequence homology to mouse/human orthologs varies and determines cross‑species utility. Exact cross‑reactivity for this item is Not specified; consult supplied sequences for alignment analyses.

Buffer Applications

Not typically applicable. This product is a nucleic acid reagent (siRNA) rather than a buffering compound.

General handling buffers (literature; not item-specific):

• Reconstitution/dilution: RNase‑free water or low‑salt Tris‑EDTA (e.g., 10 mM Tris·HCl, 0.1–1 mM EDTA, pH 7.5–8.0) to prepare stock solutions.
• Working solutions: prepare in serum‑free, antibiotic‑free medium for complexation with transfection reagents, then add to complete medium per reagent instructions.

Green Alternatives

Context: siRNA experiments are aqueous and generally avoid volatile organic solvents; environmental considerations focus on minimizing hazardous reagents and plastic waste.

Greener practice suggestions (general; not item-specific):

• Prefer aqueous formulations: reconstitute and dilute in RNase‑free water or mild buffers instead of solvent‑assisted delivery.
• Select low‑toxicity transfection reagents: some cationic lipids/polymers have improved biodegradability and lower cytotoxicity compared with older formulations; evaluate performance to potentially reduce dose.
• Optimize dosing: lower siRNA and reagent amounts to minimize waste while maintaining knockdown.
• Consolidate plastics: use multi‑channel pipetting and minimal tube changes to reduce consumable waste; recycle packaging where permitted.
• Energy efficiency: store long‑term at -20 to -80 °C but avoid unnecessary ultra‑low freezer space by aliquoting only what you need.

Comparison (literature, qualitative):

• Chemical knockdown (small‑molecule JAK inhibitors) vs genetic knockdown (siRNA): siRNA offers target specificity without introducing additional solvent burdens; small molecules may require DMSO handling and generate hazardous waste. Trade‑off: siRNA requires transfection reagents and cold storage.

Item-specific environmental attributes: Not specified for this item.

Pharmaceutical Uses

Not applicable to this catalog item. This siRNA is provided for laboratory research use only (per Product Data: For research use only) and is not formulated, labeled, or tested for any clinical or therapeutic application.

General context (informational; not item-specific):

• Oligonucleotides can serve as active pharmaceutical ingredients or excipients in regulated settings when manufactured under GMP with defined specifications and controls; this product’s grade/purity and regulatory status are Not specified for this item.
• Any use in drug development workflows (e.g., target validation assays) remains strictly preclinical research and requires appropriate quality assessment and documentation.

Physical Properties

Item-specific physical properties: Not specified for this item; refer to CoA/Spec Sheet.

General/literature values and behavior for siRNA (not item-specific):

• Physical state: dry, lyophilized solid or aqueous solution (RNase‑free). Appearance varies from white to off‑white powder or clear/colorless solution.
• Solubility: readily soluble in RNase‑free water or low‑salt buffers (e.g., 10 mM Tris·HCl, pH 7.5–8.0; 0.1–1 mM EDTA) at ≥100 µM (literature, typical practice).
• pH stability: most unmodified siRNA duplexes are stable near neutral pH; hydrolysis accelerates at alkaline pH and elevated temperature (literature).
• Tm: sequence‑dependent; A‑form RNA duplexes commonly exhibit Tm in the 50–85 °C range under standard salt (literature). Not specified for this item.
• UV absorbance: λmax ~260 nm; extinction coefficients are sequence-dependent (literature). Not specified for this item.
• Density, refractive index, logP, BP/MP: not applicable/meaningful for oligonucleotide duplexes in aqueous systems; Not specified for this item.

Practical notes:

• Prepare and handle exclusively with RNase‑free consumables to preserve integrity.
• Avoid divalent metal ions (Mg2+, Mn2+) in long‑term storage buffers unless chelated; they can catalyze RNA degradation (literature).

Quality and Grades

Item-specific grade/purity: Not specified for this item; refer to CoA/Spec Sheet.

What “pre‑designed siRNA set” generally implies (literature/generic info; not item-specific):

• Design: sequences selected by algorithms to target the specified gene (Stat6, rat) while minimizing off‑target complementarity and immunostimulatory motifs.
• Set/Pooled format: often supplied as multiple independent duplexes targeting distinct regions to increase knockdown robustness and mitigate sequence‑specific off‑targets. Number of duplexes and pooling format are Not specified for this item.
• Typical QC for siRNA vendors: MALDI‑TOF or ESI‑MS for mass confirmation, analytical HPLC purity, OD260 quantitation, residual salt/solvent assessment, and nuclease contamination checks (RNase/DNase/protease). Actual tests for this item are Not specified.
• Stabilizers/Modifications: Some siRNAs include 2′‑OMe or phosphorothioate linkages to enhance stability; presence/extent are Not specified for this item.

How to use the documentation:

• Request and review the Certificate of Analysis for: sequence(s), length, modification map, % purity by HPLC, mass spec match, and amount (nmol/OD260). These define the actionable specifications for method development and regulatory documentation in research contexts.

Reaction and Applications

Item-specific applications listed: Not specified for this item. Manufacturer Applications field is empty.

General/literature applications of Stat6-targeting siRNA in research (not item-specific):

• Gene knockdown: transient suppression of rat Stat6 mRNA and protein to interrogate IL‑4/IL‑13 signaling, Th2‑associated transcriptional programs, and JAK‑STAT pathway crosstalk in relevant cell models (e.g., primary rat immune cells, airway epithelial lines).
• Pathway mapping: define Stat6‑dependent transcription by RNA‑seq or qPCR following cytokine stimulation; validate with phospho‑STAT6 assays (e.g., pY641 Western blot).
• Phenotypic studies: assess effects on class‑switch recombination markers, mucin gene expression, or alternative macrophage activation markers in rat systems.
• Reporter assays: use STAT‑responsive luciferase reporters to quantify functional knockdown.
• Rescue experiments: re‑express siRNA‑resistant Stat6 constructs to confirm on‑target phenotype.

Practical tips (literature):

• Use a range of siRNA concentrations (e.g., 5–50 nM final) to balance efficacy and off‑target risk; determine minimal effective dose.
• Include controls: non‑targeting siRNA, positive knockdown control, and transfection‑only mock.
• Time course: measure mRNA knockdown at 24–48 h and protein at 48–96 h.
• Pooling: combining multiple duplexes targeting distinct Stat6 sites can increase robustness; specifics for this item are Not specified.

Reaction Conditions

Chemical reaction conditions are not applicable. Below are general/literature conditions for functional use of siRNA in cells (not item-specific):

• Complex formation: incubate siRNA with a cationic lipid/polymer transfection reagent in serum‑free medium for 10–30 min at room temperature, following the reagent’s specified N/P ratio.
• Dosing: typical final siRNA concentration 5–50 nM; optimize for cell type to minimize off‑targets and cytotoxicity.
• Cell density: 30–70% confluence at transfection commonly yields good uptake for adherent lines; primary cells may require electroporation or specialized reagents.
• Incubation: add complexes to cells for 4–24 h, then replace with complete medium if required by the reagent.
• Readouts: assess mRNA knockdown at 24–48 h; protein reduction at 48–96 h; functional assays thereafter.
• Controls: non‑targeting siRNA, transfection control (e.g., siGLO), and a positive knockdown control for benchmarking.

These parameters are offered as general guidance from the literature. Item‑specific recommended conditions are Not specified for this item; consult the CoA and your transfection reagent documentation.

Safety and Handling

GHS classification and hazard statements: Not specified for this item; consult the product SDS for authoritative safety information.

General handling guidance for siRNA reagents (not item-specific):

• PPE: wear lab coat, safety glasses, and appropriate gloves. Treat as a laboratory chemical/biological reagent. Avoid inhalation of powders/aerosols.
• RNase control: use RNase‑free tips, tubes, and solutions. Clean work surfaces and tools with RNase decontamination solutions. Change gloves frequently.
• Incompatibilities: avoid strong oxidizers and alkalis for solutions; avoid nucleases (RNases) from skin contact or contaminated plastics.
• First aid (general): in case of contact with eyes/skin, rinse with water. If inhaled, move to fresh air. If swallowed, rinse mouth. Seek medical attention if symptoms occur. Defer to SDS for details.
• Waste: dispose of according to institutional guidelines for non‑hazardous nucleic acid waste unless local regulations specify otherwise.

Storage and shipping (item-specific where provided):

• Storage Conditions: Store at -20°C (per Product Data). Minimize freeze‑thaw by aliquoting upon first thaw.
• Shipped In: Ice chest + Ice pads (per Product Data) to maintain cold chain.

Transportation: typically non‑dangerous goods; verify with SDS and carrier rules.

Solvent Selection

Applicability: For siRNA, the relevant “solvents” are aqueous buffers rather than organic solvents.

General/literature guidance (not item-specific):

• Primary diluent: RNase‑free water is preferred for initial reconstitution to a convenient stock (e.g., 20–100 µM). TE‑low (10 mM Tris·HCl, 0.1–1 mM EDTA, pH 7.5–8.0) can improve long‑term stability by chelating trace divalent metals.
• Avoid: repeated freeze‑thaw cycles; high ionic strength (>200 mM NaCl) during long‑term storage; strong acid/alkali. For transfection, final working buffer is dictated by the transfection reagent and cell compatibility.
• Co‑solvents: Organic co‑solvents are generally unnecessary and discouraged; even low % DMSO may affect cell physiology and transfection efficiency.

Quick comparison (literature):

• RNase‑free water: maximal versatility; best for lyophilizate reconstitution; add buffer later as needed.
• Tris‑EDTA (low EDTA): enhanced stability via metal chelation; verify compatibility with downstream enzymatic steps or transfection mixes (some cationic lipids are sensitive to EDTA or high Tris).
• PBS: acceptable for short‑term handling but not ideal for long‑term storage due to salts and divalent cations in some formulations.

Item-specific solvent recommendations: Not specified for this item; refer to CoA/Spec Sheet.

Storage and Reconstitution

Item-specific storage and shipping:

• Storage Conditions: Store at -20°C (per Product Data). Protect from repeated freeze‑thaw. Aliquot upon first thaw.
• Shipped In: Ice chest + Ice pads (per Product Data) to maintain cold chain.

Item-specific details such as supplied amount, solvent, and concentration: Not specified for this item; refer to CoA/Spec Sheet.

General/literature guidance for siRNA (not item-specific):

• Upon receipt: If lyophilized, centrifuge briefly to collect material at the bottom of the tube before opening. If in solution, keep on ice and proceed to aliquoting.
• Reconstitution: Add RNase‑free water or low‑salt TE to a convenient stock concentration (e.g., 20–100 µM). Mix gently; avoid vigorous vortexing that may shear or aerosolize.
• Aliquoting: Prepare single‑use aliquots to avoid freeze‑thaw. Use RNase‑free, low‑binding tubes and tips.
• Storage: Short term (≤1–2 weeks) at 2–8 °C; long term at -20 °C to -80 °C. Protect from light if fluorescent labels are present (labels for this item are Not specified).
• Thawing: Thaw on ice; briefly spin before opening. Keep on ice during setup.
• Handling: Work in an RNase‑controlled area. Avoid divalent cations in storage buffer; low EDTA (0.1–1 mM) can help chelate trace metals if compatible with downstream steps.

Structure and Identity

Item-specific identifiers (CAS, CID, InChIKey, SMILES): Not specified for this item; refer to CoA/Spec Sheet.

Item description (from product name; item-specific):

• Target: rat Stat6 (Signal transducer and activator of transcription 6)
• Format: Pre-designed siRNA Set A (number of duplexes, sequences, chemical modifications: Not specified for this item; refer to CoA/Spec Sheet.)

General/literature description of siRNA structure (not item-specific):

• Typical siRNA reagents are double‑stranded RNA oligonucleotides, ~19–21 bp with 2‑nt 3′ overhangs (e.g., dTdT), designed to guide RISC-mediated cleavage of complementary mRNA.
• Common features include sense/antisense strands with 5′-phosphate and 3′‑hydroxyl termini; some products may carry stabilizing modifications (e.g., 2′‑OMe, phosphorothioate), which are sequence- and vendor-specific.
• 2D description: two antiparallel RNA strands forming an A‑form helix; no defined small‑molecule ring systems; backbone of ribose-phosphate repeating units; bases (A/U/G/C) hydrogen‑bonded via Watson‑Crick pairing.

Molecular formula and molecular weight:

• Molecular Formula: Not specified for this item; sequence-dependent.
• Molecular Weight: Not specified for this item; sequence- and modification-dependent (typical single siRNA duplex ~13–15 kDa; literature, approximate).

Synthetic Utility

Classical organic “synthetic utility” is not applicable. This product is a functional genomics reagent used to silence gene expression rather than a chemical building block.

Research utility (general; not item-specific):

• Modular experimental control: siRNA against Stat6 enables targeted perturbation of a single transcription factor within complex cytokine signaling networks.
• Combination studies: pair with cytokines (IL‑4/IL‑13), JAK inhibitors, or CRISPR perturbations to dissect pathway dependencies and feedback.
• Validation workflows: use siRNA knockdown to confirm on‑target effects of small molecules or to prioritize targets for genetic knockout models.
• Epistasis and rescue: combine with overexpression of siRNA‑resistant Stat6 to establish causality.

Methodological leverage:

• Readouts include qPCR for signature transcripts (e.g., Arg1, Retnla in macrophage models; literature), Western blot for total/pY‑STAT6, reporter assays with STAT‑responsive elements, and phenotypic assays relevant to the cell type studied.

Target Specificity

Item-specific details from Product Data:

• Target gene: Stat6 (Signal transducer and activator of transcription 6)
• Species: Rat (Rattus norvegicus), per product name
• Set designation: Pre‑designed siRNA Set A

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

• Individual siRNA sequences and targeted transcript isoforms
• Number of duplexes and whether supplied as pooled or separate tubes
• Predicted off‑target analysis and genome build used for design
• Any chemical modifications (e.g., 2′‑OMe) and strand phosphorylation state

General notes (literature; not item-specific):

• Verify target transcript accession(s) to ensure coverage of the predominant Stat6 isoform in your rat strain.
• Perform BLAST alignment to screen potential off‑targets, particularly 7‑mer seed matches in 3′ UTRs.
• Use orthogonal validation (rescue with siRNA‑resistant Stat6, or multiple independent duplexes) to confirm on‑target effects.