ESYT1 Human Pre-designed siRNA Set A

Cat. No.: E1487424

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Visão geral

ESYT1 Human Pre-designed siRNA Set A contains three designed siRNAs for ESYT1 gene (Human), as well as a negative control, a positive control, and a FAM-labeled negative control. Components ESYT1 siRNA-1: 5 nmol (HPLC) ESYT1 siRNA-2: 5 nmol (HPLC) ESYT1 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

Only application detail provided in Product Data: “Pre-designed siRNA Set A” targeting human ESYT1. No tested applications, recommended dilutions, or step-by-step procedures are specified for this item.

• For execution, refer to the product CoA/Spec Sheet and follow the instructions of your chosen transfection reagent. Any additional protocols are not specified for this item.

Biological Roles

• Item-specific target

  • Gene: ESYT1 (human; Extended Synaptotagmin-1). The siRNA set is intended to downregulate ESYT1 expression in research systems.

• ESYT1 function (literature; general biology, no clinical claims)

  • ESYT1 is a membrane protein localized at endoplasmic reticulum (ER)–plasma membrane (PM) contact sites. It contains an N-terminal hydrophobic region for ER anchoring, an SMP (synaptotagmin-like, mitochondrial-lipid-binding protein) domain implicated in lipid transfer, and multiple C2 domains mediating Ca2+-dependent PM association.
  • Roles include tethering ER to PM, facilitating lipid exchange and signaling lipid organization, and responding to cytosolic Ca2+ elevations to modulate contact site dynamics.
  • ESYT family members (ESYT1/2/3) can exhibit functional redundancy; ESYT1 knockdown may be complemented or contrasted with ESYT2/3 perturbation to dissect specific roles.

• Research readouts after ESYT1 knockdown (literature examples)

  • Reduced recruitment of ESYT1 to PM upon Ca2+ influx; altered phosphoinositide distribution at the PM; potential changes in membrane repair kinetics and endocytosis/exocytosis coupling.
  • Molecular assays: qRT-PCR for ESYT1 mRNA; immunoblotting for protein levels; TIRF/confocal microscopy to visualize ER–PM contacts using fluorescent markers.

• Pathway context

  • Intersects with Ca2+ signaling, phosphoinositide metabolism, and membrane contact site biology. Interactions with E-Syts and other tethers (e.g., VAPs) form a broader ER–PM interface network.

Buffer Applications

This product is not a buffering agent. Buffer preparation guidance is not typically applicable.

• Practical note (general)
  • For dissolution and storage, siRNA is commonly handled in RNase-free water or low-salt TE buffer. Exact buffer composition for this item is not specified; refer to the CoA/Spec Sheet if buffer details are required.

Green Alternatives

While “green alternatives” are less applicable to aqueous siRNA reagents than to organic solvents, sustainability considerations can still guide experimental choices.

• Current product context

  • siRNA stocks are prepared and used in aqueous, RNase-free buffers—already favorable from a solvent green-chemistry standpoint.

• Greener practice options (general)

  • Use aqueous formulations exclusively during handling and delivery; avoid organic co-solvents unless required by specialized delivery systems.
  • Choose lipid or polymer transfection reagents with lower cytotoxicity to reduce waste from repeat dose–finding cycles.
  • Minimize plastic waste by preparing concentrated master stocks and aliquots to reduce repeated tube usage and freeze–thaw cycles.
  • Implement RNase-free, non-hazardous surface decontaminants instead of strong oxidizers where feasible.

• Alternatives to consider (contextual comparison; literature)

  • shRNA vectors (DNA-based) enable stable knockdown but require viral or plasmid delivery and generate more biological waste; siRNA is transient, aqueous, and avoids viral systems.
  • CRISPR interference (CRISPRi) provides programmable, longer-term repression but necessitates gene-editing components; siRNA offers rapid, reversible modulation with simpler aqueous workflows.

• Tradeoffs

  • siRNA provides speed and lower hazard solvent use, but requires careful RNase control; vector-based approaches may reduce reagent consumption over long timelines at the cost of added biosafety complexity.

Pharmaceutical Uses

No pharmaceutical or clinical use is indicated. For research use only (as stated in Product Data).

• General context (non-therapeutic)
  • In laboratory settings, siRNA reagents are used for target validation, pathway analysis, and assay development. They are not formulated as excipients or active pharmaceutical ingredients under this listing.
  • There is no pharmacopeial status or GMP claim for this item specified; consult the CoA if regulatory-grade information is needed.

Physical Properties

• Item-specific values

  • Appearance: Not specified for this item; refer to CoA/Spec Sheet.
  • Molecular weight (MW): Not specified for this item; refer to CoA/Spec Sheet.
  • Molecular formula: Not specified for this item; refer to CoA/Spec Sheet.
  • Solubility: Not specified for this item; refer to CoA/Spec Sheet.

• General/literature characteristics for siRNA (for planning purposes; not product specifications)

  • Typical duplex length: 19–27 bp with 2-nt 3′ overhangs.
  • Approximate MW: ~13–17 kDa per 21-mer duplex (sequence-dependent).
  • Physical state: supplied lyophilized or in aqueous buffer; readily soluble in RNase-free water or TE buffer due to polyanionic phosphate backbone.
  • Charge: highly anionic at physiological pH; does not cross membranes without a delivery reagent.
  • UV absorption: strong at 260 nm; typical extinction coefficients are sequence-dependent (nearest-neighbor calculation needed for exact E260).
  • Stability: dry and frozen solutions are stable for extended periods; aqueous solutions susceptible to RNase degradation and repeated freeze–thaw.

• Refractive index, density, pKa, logP, MP/BP

  • Not applicable/meaningful for heterogeneous biopolymers; 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 (general)

  • Design: computationally selected duplexes targeting human ESYT1 transcripts (e.g., RefSeq/Ensembl), screened to minimize off-targets and immunostimulatory motifs (details for this item are not specified).
  • Pooling: multiple siRNA duplexes combined to improve knockdown efficiency and consistency across splice variants (exact count and sequences not specified).
  • Typical QC for research-grade siRNA (literature): mass confirmation by MALDI-TOF/ESI-MS, desalting or cartridge purification, and OD260-based quantitation. HPLC purification and endotoxin testing may be provided by some vendors but are not specified for this item.

• Implications for experimental use

  • Pooled siRNA can reduce dose requirements and mitigate sequence-specific artifacts; however, optimization of transfection conditions and dose–response curves remains essential.
  • For sensitive applications (e.g., primary cells), consider endotoxin and dsRNA impurity levels; these parameters are not specified for this item and should be confirmed on the CoA if critical.

Reaction and Applications

• Item-specific applications (from catalog): Not specified beyond being a “Pre-designed siRNA Set A” targeting human ESYT1.

• General applications for siRNA targeting ESYT1 (literature; no medical/clinical claims)

  • Transient gene knockdown of ESYT1 in human cell lines to study ER–plasma membrane contact sites, lipid transfer, and Ca2+-regulated membrane tethering.
  • Pathway interrogation: assess effects on phospholipid homeostasis, PI(4,5)P2 dynamics, store-operated Ca2+ entry interfaces, and organelle contact site biology.
  • Phenotypic assays: changes in cell spreading, membrane repair, or endocytosis rates upon ESYT1 depletion; combinatorial knockdown with ESYT2/3 for redundancy studies.
  • Validation workflows: siRNA-mediated knockdown followed by qRT-PCR for mRNA reduction and immunoblotting or microscopy for protein level/localization changes.

• Practical tips (general)

  • Optimize transfection reagent, siRNA dose (e.g., low to mid-nanomolar), and cell density for each line; include non-targeting and positive-control siRNAs.
  • Time course: mRNA reduction typically detectable within 24–48 h; maximal protein reduction often at 48–96 h depending on protein half-life.
  • Minimize off-targets by using pooled siRNA and maintaining the lowest effective concentration; confirm on-target effects with at least two independent duplexes when possible (Set A may already provide pooling; specifics not provided).

Reaction Conditions

Classical reaction conditions (solvent, temperature, catalysts) are not applicable. For experimental planning, the following literature-based transfection parameters may be useful (not product specifications):

• Typical transfection setup (literature, varies by cell line and reagent)

  • Cell density at transfection: ~40–70% confluence for adherent lines to balance uptake and viability.
  • siRNA working concentration: often 5–50 nM; optimize to the lowest effective dose.
  • Complexation: dilute siRNA and transfection reagent separately in serum-free medium, incubate ~5–20 min after mixing; add to cells for 4–24 h before medium change (reagent-dependent).
  • Knockdown timeline: mRNA reduction at 24–48 h; protein nadir at 48–96 h.

• Controls and validation (general)

  • Include non-targeting siRNA controls, a toxicity control (reagent alone), and a positive-knockdown control targeting a housekeeping gene with known phenotype (where appropriate).
  • Quantify silencing by qRT-PCR and protein by immunoblot or immunofluorescence; rescue with siRNA-resistant constructs strengthens causality.

• Notes

  • Primary cells or delicate lines may require lower doses and alternative delivery (electroporation, nanoparticles). Always consult the specific transfection reagent instructions.

Safety and Handling

• GHS/SDS information (item-specific)

  • Signal word: Not specified for this item; refer to SDS.
  • Hazard statements (H-Statements): Not specified for this item; refer to SDS.
  • GHS classification and pictograms: Not specified for this item; refer to SDS.

• General handling guidance for siRNA oligonucleotides (literature/good practice)

  • PPE: laboratory coat, gloves (nitrile), and safety glasses. Change gloves frequently to avoid RNase contamination.
  • RNase control: use RNase-free consumables and solutions; clean work surfaces with RNase decontamination solutions; avoid touching tube rims/caps.
  • Incompatibilities: avoid strong acids/bases and nucleases; avoid prolonged exposure to elevated temperature. Do not mix with divalent cation–rich buffers without appropriate complexing agents if transfection is intended.
  • First aid (overview; defer to SDS): in case of inhalation of aerosolized material, move to fresh air; after skin/eye contact, rinse with water; if ingested, rinse mouth and seek medical advice.
  • Environmental considerations: dispose of oligonucleotide-containing waste according to institutional biosafety guidelines; although non-infectious, treat as bio-research waste.

• Special risks

  • siRNA is not known to be volatile or flammable; primary risk is biological cross-contamination (RNases) and experimental exposure when aerosolizing solutions during pipetting.

Solvent Selection

This product is an siRNA reagent and is typically used in aqueous, RNase-free media rather than organic solvents.

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

• General guidance (literature)

  • Primary solvents: RNase-free water or TE buffer (10 mM Tris-HCl, 0.1–1 mM EDTA, pH ~7.5–8.0) for stock solutions; low-salt buffers recommended to maintain duplex integrity.
  • Working diluent: serum-free, antibiotic-free cell culture medium during complexation with transfection reagents; complete growth medium after complex formation.
  • Miscibility: siRNA is water-soluble; insoluble in nonpolar organic solvents. Alcohols (e.g., ethanol, isopropanol) are used for precipitation, not dissolution.
  • Avoid: solutions containing RNases or high concentrations of divalent cations prior to complexation; extremes of pH that can promote hydrolysis.

• Comparison (general)

  • RNase-free water vs TE: TE offers improved nuclease protection (via EDTA chelation) for long-term storage; water is suitable for short-term use and when EDTA would interfere with downstream steps.

Storage and Reconstitution

• Item-specific storage and shipping (from Product Data)

  • Storage: Store at -20°C
  • Shipped in: Ice chest + ice pads

• Reconstitution and handling

  • Reconstitution medium, target concentration, and buffer composition: Not specified for this item; refer to CoA/Spec Sheet.

• General guidance (literature; not product specifications)

  • Upon receipt, briefly centrifuge vials to collect contents. Reconstitute in RNase-free water or TE buffer to a convenient stock (e.g., 10–50 µM), aliquot to avoid repeated freeze–thaw, and store at -20°C or below. For long-term storage, -80°C is commonly used.
  • Avoid multiple freeze–thaw cycles; thaw on ice and mix gently. Maintain RNase-free technique throughout.

• Stability note

  • Lyophilized siRNA is typically stable at -20°C when protected from moisture. Aqueous stocks are generally stable for months at -20°C with EDTA-containing buffer, but exact stability for this item is not specified; consult CoA/Spec Sheet.

Structure and Identity

Brief description: ESYT1 Human Pre-designed siRNA Set A is a pool of short interfering RNA (siRNA) duplexes designed to silence the human ESYT1 gene via RNA interference (RNAi).

• Item-specific facts (from Product Data)

  • Product type: Pre-designed siRNA set targeting human ESYT1 (Extended Synaptotagmin-1)
  • Format: siRNA duplexes (exact sequences, lengths, and number of duplexes in Set A are not specified)
  • CAS: Not specified for this item; refer to CoA/Spec Sheet.
  • 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 / InChI / InChIKey: Not applicable for heterogeneous oligonucleotide mixtures; not specified for this item.

• General structural description (literature)

  • siRNA typically consists of a double-stranded RNA helix ~19–27 base pairs with 2-nt 3′ overhangs, composed of ribonucleotides linked by 3′→5′ phosphodiester or phosphorothioate backbones.
  • Functional groups: multiple phosphate groups (polyanionic), ribose sugars bearing 2′-hydroxyls (unless chemically modified), and nucleobases (A, U, G, C; sequence-specific).
  • 2D/secondary structure: anti-parallel duplex with Watson–Crick base pairing; may include terminal overhangs; no defined stereocenters beyond the natural ribose configuration.

• Notes on design (general)

  • Pre-designed pools commonly include multiple non-overlapping siRNA duplexes to improve knockdown robustness and reduce off-target risk; exact composition for this item is not specified.

Synthetic Utility

Not a chemical building block for synthetic transformations. As an siRNA reagent, its value lies in functional genomics rather than in classical organic synthesis.

• Relevant utility (general research)
  • Enables target validation and phenotypic screening by transient gene knockdown of ESYT1, informing upstream/downstream pathway mapping that can guide later small-molecule design.
  • Can be combined with chemical perturbations (e.g., lipid metabolism inhibitors) to perform epistasis-like experiments that inform synthetic and medicinal chemistry prioritization.

Target Specificity

• Item-specific information (from Product Data)
  • Target gene: ESYT1 (human)
  • Reagent type: Pre-designed siRNA Set A (pool of siRNA duplexes)
  • Additional details (sequences, number of duplexes, transcript isoform coverage, off-target screening): Not specified for this item; refer to CoA/Spec Sheet.