ANKAR Human Pre-designed siRNA Set A

Cat. No.: A1466356

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Visão geral

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

biological_products

Application Protocols

Item-specific, tested application protocols are not provided in the Product Data.

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

General reminder

• Follow your transfection reagent manufacturer’s protocol for siRNA delivery and optimize dose, cell density, and exposure time per cell line. Include appropriate controls (non-targeting, positive control, and mock).

Biological Roles

Item-specific target

• Target designated by product name: human ANKAR. No sequence, transcript IDs, or functional annotations are provided in the Product Data.

General context (literature)

• Ankyrin repeat–containing proteins: Ankyrin repeats are common protein–protein interaction motifs found in diverse regulators, scaffolds, and signaling adaptors. Proteins harboring these motifs often participate in assembly of multi-protein complexes and modulation of signaling pathways.
• Gene silencing utility: siRNA-mediated reduction of a target gene’s mRNA enables interrogation of its role in cellular processes, including effects on signaling, trafficking, or structural organization, depending on the gene’s function.
• Verification: Because gene families with ankyrin repeats can have paralogs with related domains, confirm specificity via multiple independent siRNAs and rescue experiments to attribute phenotypes to ANKAR specifically.

Caveat

• Specific cellular functions, pathways, or phenotypes associated with the human ANKAR gene are not provided here and should be derived from primary literature and databases (UniProt, NCBI Gene, Ensembl) prior to experimental planning.

Research use

• For research use only. Not for diagnostic or therapeutic applications.

Buffer Applications

This product is not a buffering agent and is not used to prepare classical laboratory buffer systems.

Practical note (literature)

• Reconstitution and working solutions are typically prepared in RNase-free water or low-ionic strength buffers (e.g., 10 mM Tris·HCl, 0.1–1 mM EDTA, pH ~7.5–8.0) that help maintain siRNA integrity. For complexation with transfection reagents, follow the reagent’s recommended diluent (often serum-free medium such as Opti-MEM).

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

Green Alternatives

Context

• As an aqueous, non-volatile oligonucleotide reagent, the primary environmental considerations stem from ancillary materials (transfection reagents, plastics, and buffers), not the siRNA itself.

Greener practice suggestions (literature)

• Reduce single-use plastics: Consolidate aliquots where feasible, employ low-bind but recyclable plastics, and plan experiments to minimize consumables.
• Transfection reagents: Select lower-toxicity, serum-compatible reagents that enable reduced medium exchanges, lowering waste.
• Solvent use: Avoid organic solvents for siRNA handling; prepare and dilute in water or buffered saline. If ethanol is required for sterilization, use minimal volumes and allow full evaporation.
• Energy footprint: Store working aliquots at −20°C and bulk at −80°C only if necessary; avoid unnecessary ultra-cold storage. Maintain efficient cold-chain logistics to reduce dry ice usage.
• Waste and decontamination: Prefer commercial RNase decontaminants over DEPC where appropriate to avoid hazardous waste; if DEPC is used, neutralize and dispose per regulations.

Comparison (qualitative)

• siRNA pools vs plasmid shRNA: Transient siRNA avoids viral packaging and antibiotic selection, reducing biosafety burden and waste, at the cost of repeated dosing. Choice depends on experimental duration and resources.

Pharmaceutical Uses

This is a research-grade siRNA reagent.

Item-specific regulatory/compendial status

• Grade/purity and pharmacopeial listings: Not specified for this item; refer to CoA/Spec Sheet.

General context (literature)

• Role in development workflows: siRNAs are widely used in discovery biology, target validation, and preclinical mechanism-of-action studies within pharmaceutical research labs. They enable rapid, reversible gene suppression without genomic modification.
• Formulation analogs: While therapeutic siRNA formulations exist in the literature (e.g., with lipid nanoparticles), those are distinct GMP-grade products with stringent quality, stability, and sterility requirements that are not applicable to this research reagent.
• Handling in regulated settings: If used in GLP research, document lot-specific identity, purity, and analytical data (sequence verification, UV quantitation) as provided by the manufacturer.

Restriction

• For research use only. Not for use in humans or clinical diagnostics.

Physical Properties

Item-specific facts (from Product Data)

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
• Other physico-chemical specs (MP/BP, density, UV cutoff, RI): Not applicable/not specified for this item.

General properties (literature)

• Physical state: Typically supplied lyophilized or in aqueous RNase-free buffer; forms clear solution upon reconstitution.
• Solubility: Freely soluble in RNase-free water or low-salt buffers (e.g., 1× siRNA buffer, 10 mM Tris·HCl, 1 mM EDTA, pH ~7.5–8.0). Insoluble in most organic solvents.
• UV characteristics: Strong absorption at 260 nm (A260). Extinction coefficient depends on exact sequence and length; calculate from sequence if provided.
• Charge/ionization: Strongly polyanionic at neutral pH due to phosphate backbone; counterions (e.g., Na+, K+) influence hygroscopicity.
• Stability (qualitative): siRNAs are chemically stable when dry and stored frozen; hydrolysis and RNase-mediated degradation occur in aqueous/biological environments. Avoid repeated freeze–thaw.
• Typical duplex size (literature): 19–21 bp core with 2-nt 3′ overhangs; approximate MW per 21mer duplex often in the 13–14 kDa range, but exact value is sequence-dependent and not specified for this item.

Quality and Grades

Item-specific facts (from Product Data)

• Grade/purity and purification method: Not specified for this item; refer to CoA/Spec Sheet.

General considerations for siRNA quality (literature)

• Purification level: siRNAs may be supplied as desalted, cartridge (PAGE) purified, or HPLC-purified. HPLC/PAGE reduces shortmers and ensures tighter length distribution, which can improve potency and reduce off-targets.
• Identity/verification: Typical QC includes MALDI-TOF/ESI-MS of single strands, UV A260 quantitation, and OD260-to-mass conversion using strand-specific extinction coefficients. For pools, vendors often mix equimolar duplexes.
• Residual salts/solvents: Low-salt content is preferred for transfection. Any stabilizers or carrier nucleic acids (e.g., yeast tRNA) should be disclosed on the CoA.
• End-modifications: 5′-phosphate on antisense strand and 2′-O-methyl or phosphorothioate modifications at select positions can enhance stability and reduce immunostimulation; presence/absence should be confirmed on the Spec Sheet (not specified for this item).
• Functional QC: Knockdown performance is often characterized in a reference cell line at 10–50 nM with RT-qPCR/Western readouts; consult the manufacturer’s data when available (not provided for this item).

Reaction and Applications

Focus for this reagent is RNA interference (RNAi) in mammalian systems rather than chemical synthesis.

Applications (literature)

• Gene silencing: Sequence-specific knockdown of human ANKAR transcripts by guiding Argonaute-containing RISC to complementary mRNA, promoting endonucleolytic cleavage and degradation.
• Pooled design rationale: Using multiple siRNAs targeting non-overlapping regions of ANKAR can improve knockdown robustness and mitigate single-sequence off-target artifacts.
• Functional genomics: Use in pathway mapping, target validation, and phenotypic screening where reduction of ANKAR expression is desired.
• Controls: Employ non-targeting siRNA controls, positive silencing controls (e.g., PLK1), and rescue experiments (siRNA-resistant cDNA) to confirm on-target effects.

Practical tips (literature)

• Transfection optimization: Titrate siRNA (e.g., 5–50 nM) and reagent amount per cell type. Optimize cell density and serum conditions.
• Timing: Maximal mRNA reduction typically observed at 24–48 h; protein knockdown may peak at 48–96 h depending on protein half-life.
• Off-target minimization: Keep siRNA dose low yet effective; consider pooling and chemical modifications; validate by at least two independent siRNAs.
• Verification: Quantify knockdown by RT-qPCR and/or immunoblotting; monitor cell health and morphology.

Item-specific application claims or performance data: Not specified for this item; refer to CoA/Spec Sheet.

Reaction Conditions

Chemical reaction conditions are not applicable. For experimental planning, the following literature-based RNAi conditions are provided as general guidance (not item-specific):

General RNAi conditions (literature)

• Working concentration: Commonly 5–50 nM siRNA final in cell culture; optimal concentration depends on cell type and transfection reagent.
• Complexation: Mix siRNA with a compatible transfection reagent in serum-free medium; incubate 5–30 min to form lipoplexes/polyplexes per reagent instructions.
• Cell density: Plate cells to be 40–70% confluent at transfection to balance viability and uptake.
• Incubation: Replace with complete medium after 4–24 h if required by the reagent; peak mRNA knockdown at ~24–48 h, protein knockdown at ~48–96 h, depending on protein half-life.
• Readouts: RT-qPCR for mRNA, Western blot/ELISA/IF for protein. Include non-targeting and positive control siRNAs.

Item-specific recommendations (buffers, concentrations, sequences, validated cell lines, expected knockdown): Not specified for this item; refer to CoA/Spec Sheet.

Safety and Handling

Item-specific facts (from Product Data)

• GHS classification, pictograms, H-statements, signal word: Not specified for this item; refer to SDS.
• Storage: Store at −20°C. Shipped on ice packs in an insulated container.

General guidance (literature/SDS best practices)

• Expected hazards: Synthetic siRNAs are generally considered low hazard; however, dusts/aerosols should be avoided. Handle as a laboratory chemical reagent.
• PPE: Wear lab coat, gloves (nitrile), and safety glasses. Use RNase-free consumables and change gloves frequently to avoid RNase contamination.
• Engineering controls: Work in a clean area; for solution prep, consider a designated RNase-free hood or PCR workstation. Avoid generating aerosols.
• Incompatibilities: RNases (ubiquitous on skin/surfaces) rapidly degrade siRNA; avoid contaminated buffers and plastics. Strong acids/bases and elevated temperatures accelerate hydrolysis.
• First aid (overview): If inhaled—move to fresh air. Skin/eye contact—rinse with water. If ingested—rinse mouth. Seek medical attention if symptoms persist. Follow institutional SDS procedures.
• Waste: Dispose of aqueous nucleic acid waste per local regulations. Decontaminate surfaces with RNase deactivation solutions (e.g., freshly prepared 0.1% DEPC for equipment as appropriate) or commercial RNase removers.
• Transport/stability: Maintain cold chain as provided on receipt; prolonged exposure to ambient temperatures may affect long-term stability after reconstitution.

Solvent Selection

Applicability

• This product is an RNA oligonucleotide set intended for aqueous reconstitution; organic solvent selection is not typically applicable.

Practical guidance (literature)

• Primary diluents: Use nuclease-free water, 1× siRNA buffer, or low-ionic strength TE (10 mM Tris·HCl, 0.1–1 mM EDTA, pH 7.5–8.0) to enhance stability and minimize divalent-cation–catalyzed hydrolysis.
• Avoid: Non-aqueous solvents (e.g., alcohols, DMF, DMSO) for stock preparation, as they can affect duplex formation and downstream transfection. Small volumes of DMSO may be tolerated within some transfection mixes but are generally unnecessary.
• Compatibility: siRNA stocks are compatible with most cationic and polymeric transfection reagents when prepared in their recommended complexation buffers (e.g., Opti-MEM or serum-free media just prior to transfection).
• Concentration planning: Prepare high-concentration stocks (e.g., 10–100 µM; literature) to minimize freeze–thaw and dilution steps; filter sterilize if required using 0.22 µm RNase-free filters.
• Container materials: Use certified RNase-free polypropylene microtubes. Avoid glass for long-term storage due to potential adsorption/desorption and alkaline surface-catalyzed cleavage.

Storage and Reconstitution

Item-specific facts (from Product Data)

• Storage: Store at −20°C.
• Shipping: Shipped in an ice chest with ice pads to maintain cold chain.

Not specified for this item

• Supplied form (lyophilized vs solution), stabilizers, buffer composition, and initial concentration: Not specified for this item; refer to CoA/Spec Sheet.

General guidance (literature)

• Upon receipt: Briefly centrifuge to collect material. If lyophilized, allow vial to equilibrate to room temperature before opening to prevent condensation.
• Reconstitution: Use nuclease-free water or appropriate buffer (e.g., 10 mM Tris·HCl, 0.1–1 mM EDTA, pH 7.5–8.0). Typical stock concentrations are 10–100 µM; exact concentration should be selected based on experimental design. Label as general guidance only.
• Aliquoting: Prepare single-use aliquots in RNase-free tubes to avoid repeated freeze–thaw. Overlay with sterile RNase-free water to minimize headspace if stored in frost-free freezers.
• Storage after reconstitution: Store at −20°C (short to mid-term). For extended storage, −80°C is commonly used. Avoid >3–4 freeze–thaw cycles.
• Handling precautions: Use RNase-free tips/tubes; wear clean gloves; wipe surfaces with RNase decontaminant. Avoid prolonged exposure at room temperature.

For research use only.

Structure and Identity

Item-specific facts (from Product Data)

• Product type: Pre-designed siRNA set (Set A)
• Target: Human ANKAR (gene symbol per product name). No sequences provided.
• CAS/CID/InChIKey/SMILES: Not applicable to this oligonucleotide mixture; not specified for this item.
• Molecular formula/MW: Not specified for this item; refer to CoA/Spec Sheet.

General description (literature)

• Composition: Pool of double-stranded small interfering RNAs (siRNAs), typically 19–21 bp duplexes with 2-nt 3′ overhangs; sense and antisense strands with phosphodiester backbones and standard RNA ribose (2′-OH) unless otherwise indicated by manufacturer.
• Structural features: Each duplex comprises complementary RNA strands forming an A-form helix; typical 3′ overhangs (e.g., dTdT) and 5′-phosphate on antisense are common design features to enhance RISC loading (design varies by supplier).
• 2D/functional view in words: Linear polyanionic biopolymer (RNA) with repeating ribose–phosphate backbone and nucleobases (A, U, G, C). Duplexes align antiparallel; antisense strand guides Argonaute to complementary ANKAR mRNA.
• Form: Supplied as a mixture of multiple siRNA duplexes targeting distinct regions of the ANKAR transcript to enhance knockdown robustness and reduce off-target effects (typical for “set” products).

Synthetic Utility

Not typically applicable.

Rationale

• This product is a pool of siRNA duplexes intended for biological gene-silencing experiments. It is not a building block or reagent for chemical synthesis in the traditional sense.

Related laboratory utility (literature)

• Functional genomics tool: Enables perturbation of ANKAR expression to study pathway dependencies and genetic interactions.
• Screening: Useful in RNAi screens to identify phenotypes linked to ANKAR suppression or to evaluate compound–gene interactions (chemical–genetic synergy/antagonism).

If chemical derivatization or conjugation (e.g., dyes, peptides) is desired, consult the manufacturer; modification status for this item is not specified and should not be assumed.

Target Specificity

From Product Data

• Target gene: ANKAR (human).
• Format: Pre-designed siRNA Set A (pool of multiple siRNA duplexes targeting the same gene).

Not provided in Product Data

• Exact siRNA sequences, target transcript IDs/isoforms, off-target assessment, BLAST homology, and cross-species reactivity: Not specified for this item; refer to CoA/Spec Sheet.

General notes (literature)

• Pooled siRNA sets are typically designed to minimize off-target effects by diluting any one sequence’s unintended interactions while maintaining on-target potency.
• Specificity should be validated experimentally via multiple orthogonal assays and, where possible, rescue with an siRNA-resistant ANKAR construct.