AHI1 Human Pre-designed siRNA Set A

Cat. No.: A1487082

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Panoramica

AHI1 Human Pre-designed siRNA Set A contains three designed siRNAs for AHI1 gene (Human), as well as a negative control, a positive control, and a FAM-labeled negative control. Components AHI1 siRNA-1: 5 nmol (HPLC) AHI1 siRNA-2: 5 nmol (HPLC) AHI1 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.

Product Line

Prodotti biologici

Application Protocols

Item-specific tested applications, recommended conditions, and detailed protocols: Not specified for this item; refer to CoA/Spec Sheet.

Note: For general transfection and knockdown workflows with siRNA, consult your transfection reagent manufacturer and institutional protocols. Follow RNase‑free technique and include appropriate controls.

Biological Roles

Item-specific target context:

• Target gene: AHI1 (Abelson helper integration site 1), Homo sapiens. This siRNA set is intended to reduce AHI1 mRNA levels in human cells (research use only).

General biology of AHI1 (literature/general):

• AHI1 encodes a scaffold protein containing WD40 repeats and an SH3 domain, implicated in assembly of protein complexes at primary cilia and in neurons.
• Cellular processes: ciliogenesis, endocytic/vesicular trafficking, and modulation of signaling hubs localized to the ciliary base or axoneme. Interactions with ciliary components can influence pathway responsiveness.
• Expression: observed in multiple tissues with enrichment in neuronal lineages and ciliated epithelia (literature patterns are tissue‑ and context‑dependent).
• Experimental readouts after AHI1 knockdown: alterations in ciliary morphology/number, changes in localization of ciliary receptors, and transcriptional signatures of cilium‑related pathways. Protein‑level assessments are typically via immunoblotting or immunofluorescence using validated anti‑AHI1 antibodies.

Notes:

• The above reflects general biological knowledge and should guide hypothesis generation for cell‑based studies. It does not constitute clinical or diagnostic guidance.
• For on‑target confirmation, pair siRNA experiments with rescue using siRNA‑resistant AHI1 constructs, and include non‑targeting siRNA controls.

Buffer Applications

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

General buffer guidance for siRNA handling (literature/general):

• Reconstitution: RNase‑free water or 10 mM Tris‑HCl, pH 7.5–8.0. For long‑term storage, TE (10 mM Tris‑HCl, 0.1–1 mM EDTA, pH 7.5–8.0) can chelate Mg²⁺ and stabilize against RNases. Verify EDTA compatibility with downstream transfection chemistry.
• Working stocks: 10–50 µM; dilute into serum‑free transfection medium (e.g., Opti‑MEM or equivalent) during complex formation per reagent guidance.
• pH and ionic strength: maintain near‑neutral pH; moderate ionic strength (50–100 mM NaCl) supports duplex stability but may affect complexation—optimize empirically.
• Filtration: if sterile solutions are required, filter through 0.22 µm low‑binding membranes under RNase‑free conditions; avoid shear from excessive plunging.
• Storage aliquots: prepare single‑use aliquots to avoid freeze–thaw cycles; label with concentration, buffer, and date.

Not typically applicable buffers:

• Classical biochemical buffer systems for enzymology or electrophoresis (e.g., HEPES, MOPS, MES) are not intrinsically required for siRNA storage; they may be used in downstream assays but should be verified for compatibility with transfection complexes.

Green Alternatives

Applicability note: siRNA use is predominantly aqueous and low‑temperature; environmental considerations focus on solvent minimization, cold‑chain logistics, and waste reduction.

Greener practice recommendations (general):

• Aqueous media: prepare and store stocks in RNase‑free water or Tris buffers—no organic solvents required.
• Minimize plastics: employ reusable, RNase‑decontaminated racks and reservoir systems; consolidate transfections to reduce consumables.
• Low‑toxicity transfection reagents: select formulations with improved biodegradability and lower cytotoxicity; validate performance equivalence.
• Energy use: limit freeze–thaw and consolidate shipments; consider lyophilized formats to reduce cold‑chain intensity when available (format for this item not specified).
• Decontamination: use non‑volatile RNase inactivators where feasible instead of DEPC (which is hazardous); if DEPC‑treatment is necessary, handle under a fume hood and neutralize properly.

Comparison (general considerations):

• DEPC‑treated water vs certified RNase‑free water: certified RNase‑free avoids handling a carcinogenic alkylating agent (DEPC), reducing hazard without compromising performance.
• Organic solvent‑based delivery (e.g., some polymer solutions) vs aqueous lipid formulations: aqueous systems reduce VOCs; ensure equal transfection efficiency before switching.

Item-specific environmental credentials or certifications: Not specified for this item; refer to CoA/Spec Sheet.

Pharmaceutical Uses

Item-specific pharmacopeial/formulation status: Not specified for this item; refer to CoA/Spec Sheet.

Context (general; no therapeutic claims):

• Pre‑designed research‑grade siRNA such as this product are intended for laboratory studies (in vitro/in cellulo). They are not manufactured under GMP and are not intended for human or veterinary use.
• In pharmaceutical R&D settings, research‑grade siRNAs are used to validate targets, explore mechanism of action, and deconvolute pathways prior to medicinal chemistry or biologics campaigns.
• Excipient considerations (general): for formulated siRNA drug substances (separate from this research item), buffers such as phosphate, citrate, or acetate and stabilizers like sugars or PEGylated lipids may be employed. Such formulation details do not apply to this item.

Compliance note:

• This product is for research use only. Any use in manufacturing or diagnostics would require appropriate quality and regulatory qualifications not described here.

Physical Properties

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

General/literature properties for siRNA (for context only):

• Physical state: lyophilized solid or aqueous solution, depending on packaging; hygroscopic; readily water‑soluble.
• Solubility: freely soluble in RNase‑free water and low‑salt Tris buffers (e.g., TE); insoluble in nonpolar organic solvents.
• Approximate molecular weight: individual 21‑mer duplexes typically 13–15 kDa, depending on base composition and modifications (literature).
• pKa: nucleobases have multiple pKa values; duplex behavior is dominated by phosphate acidity (pKa1 ~1–2; literature); functional pH range for handling is near neutral.
• UV absorbance: strong at 260 nm; typical duplex extinction coefficients 200,000–300,000 M⁻¹ cm⁻¹ for 21‑mers (literature). Item‑specific ε260 not specified.
• Melting temperature (Tm): sequence‑dependent; many 21‑bp siRNA duplexes display Tm 60–80°C in standard buffers (literature). Item‑specific Tm not specified.
• Stability: stable when dry and protected from moisture; in solution, stability increases at pH ~7–8 and low temperature; repeated freeze–thaw should be minimized.

Note: Do not treat the above as specifications. Consult the item’s CoA/Spec Sheet for any measured values associated with this SKU.

Quality and Grades

Item-specific quality details:

• Grade/Purity: Not specified for this item; refer to CoA/Spec Sheet.
• Appearance: Not specified for this item; refer to CoA/Spec Sheet.

General context for siRNA quality (literature/general):

• Common grades: desalting grade (crude, desalinated), PAGE‑purified, or HPLC‑purified. Higher‑stringency purifications reduce shortmers and n‑1 species, improving knockdown consistency and lowering off‑target risk.
• Analytical characterization: mass confirmation by MALDI‑TOF/ESI, capillary electrophoresis or ion‑pair RP‑HPLC profiles, and UV quantitation at 260 nm. Endotoxin testing may be reported for sensitive cell systems.
• Modifications: chemical stabilizers (e.g., 2′‑O‑methyl, 2′‑F, phosphorothioate, or terminal conjugates) can improve nuclease resistance and uptake; none are specified for this item.
• Sequencing disclosure: pre‑designed sets often ship with sequence IDs and target coordinates provided on the CoA; exact sequences for this SKU are not provided here.

How to use quality information:

• For demanding applications (primary cells, low‑dose transfection), prioritize HPLC/PAGE‑purified siRNA and request endotoxin levels.
• Confirm delivered amount (OD260 units or nmol) and purity metrics on the CoA before experiment planning.
• If performing qPCR or RNA‑seq readouts, ensure strand bias and passenger‑strand content are minimized; consider asymmetric designs, but this item’s design details are not specified.

Reaction and Applications

Item-specific applications provided by manufacturer: Not specified for this item; refer to CoA/Spec Sheet.

General applications for pre‑designed siRNA to human AHI1 (literature/general):

• Gene silencing: sequence‑specific knockdown of AHI1 mRNA via the RNA interference pathway. The antisense (guide) strand loads into RISC (Ago2), directing cleavage or translational repression of complementary AHI1 transcripts.
• Functional genomics: dissect AHI1’s role in ciliogenesis, vesicular trafficking, and signal scaffolding in human cell models; supports pathway mapping and synthetic‑lethality screens when combined with small molecules/CRISPR controls.
• Phenotypic assays: assess effects on ciliary markers, downstream signaling readouts, or transcriptomic changes by qPCR/RNA‑seq. Include non‑targeting and positive‑control siRNAs.
• Rescue validation: perform siRNA‑resistant cDNA rescue (silent mutations in the siRNA target site) to confirm on‑target effects.
• Practical tips (general):
  • Use 10–30 nM final siRNA as a starting range; optimize by cell type and transfection reagent.
  • Verify knockdown 24–72 h post‑transfection at mRNA level (qPCR) and 48–120 h at protein level (WB/IF), depending on protein turnover.
  • Pooling multiple siRNAs against AHI1 can mitigate off‑target effects at lower per‑strand doses; composition of this Set A is not specified for this item.
  • Implement strand‑specific RT‑qPCR or 5′ RACE for cleavage validation, where applicable.

Reaction Conditions

Not a chemical reaction reagent: classical synthetic reaction conditions do not apply.

General experimental conditions for siRNA transfection (literature/general):

• Cell lines: begin optimization in robust human lines (e.g., HEK293, HeLa, U2OS) before transferring to primary cells; use RNase‑free plastics throughout.
• Dosing: 5–50 nM siRNA final; typical starting point 10–30 nM. Lower per‑strand dose if using a pooled set. This item’s pool composition is not specified.
• Complex formation: incubate siRNA with a cationic lipid/polymer in serum‑free medium for 10–30 min at room temperature; add to cells at 40–70% confluence; replace with complete medium after 4–24 h per reagent guidance.
• Timing: assay mRNA knockdown at 24–72 h; protein knockdown may require 48–120 h depending on turnover.
• Controls: non‑targeting siRNA, transfection reagent only, and a positive control siRNA (e.g., targeting a housekeeping gene) for process verification.
• Readouts: RT‑qPCR (ΔΔCt), Western blot, immunofluorescence, or functional assays relevant to AHI1 biology.

These are general literature guidelines; always follow your transfection reagent’s instructions. Item‑specific recommended conditions are not provided.

Safety and Handling

Item-specific hazard information:

• GHS Classification, Signal Word, H‑Statements, Pictograms: Not specified for this item; refer to SDS.
• Research use only.

General safety guidance for siRNA and oligonucleotides (literature/general):

• Hazard profile: synthetic siRNA duplexes are generally considered low hazard; handle as laboratory chemicals using standard precautions. Avoid inhalation of powders and aerosol generation during reconstitution.
• PPE: lab coat, safety glasses, and disposable nitrile gloves. Change gloves frequently to maintain RNase‑free conditions.
• RNase control: use dedicated RNase‑free consumables; clean benches and pipettes with RNase decontamination solutions; work quickly on ice for solutions.
• Incompatibilities: avoid strong acids/bases and nucleases; do not mix with biocides that can modify nucleic acids unless validated (e.g., avoid residual bleach in vessels; rinse thoroughly).
• First aid (general):
  • Inhalation: move to fresh air; seek medical attention if symptoms occur.
  • Skin/eye contact: rinse with water for several minutes; remove contaminated clothing; seek medical advice if irritation persists.
  • Ingestion: rinse mouth; seek medical attention if unwell.
• Waste: dispose of aqueous nucleic‑acid waste as nonhazardous biological/chemical waste per institutional policy; decontaminate RNases via appropriate solutions if required.

Always consult the product‑specific SDS for authoritative hazard and handling information.

Solvent Selection

Item-specific instructions: None specified beyond storage at −20°C.

General guidance for siRNA (literature/general):

• Primary solvent: RNase‑free water is the default for reconstitution. Low‑salt buffers (e.g., 10 mM Tris‑HCl, pH 7.5–8.0; 0.1–1 mM EDTA [TE]) are commonly used to improve long‑term stability by chelating divalent RNase cofactors.
• Avoid: nonpolar organic solvents (insoluble), strong acids/bases, and buffers containing RNases or high ionic strength if subsequent complexation with transfection reagents is required.
• Working solutions: prepare at 10–50 µM for convenient pipetting; dilute into serum‑free transfection medium immediately before complex formation with cationic lipids or polymers.
• pH considerations: neutral to slightly basic pH (7.0–8.0) maintains ribose/phosphate integrity and duplex stability.
• Ionic strength: modest salt (e.g., 50–100 mM NaCl) can stabilize duplexes; however, complex formation with some transfection reagents is optimal in low‑ionic media—follow the reagent vendor’s guidance.

Quick comparison (literature):

• RNase‑free water: maximal compatibility; best for short‑term working stocks; lowest buffering.
• 10 mM Tris (pH 7.5–8.0): improved pH control; good for freezer stocks.
• TE (10 mM Tris, 0.1–1 mM EDTA): enhanced nuclease protection; verify EDTA compatibility with downstream transfection chemistry.

Storage and Reconstitution

Item-specific instructions (from Product Data):

• Storage: Store at −20°C.
• Shipping: Shipped in ice chest with ice pads.

Additional practical guidance (general; not item‑specific specifications):

• Upon receipt: if lyophilized, centrifuge briefly to collect material before opening. If in solution, verify concentration and buffer on the CoA.
• Reconstitution (general): dissolve in RNase‑free water or 10 mM Tris‑HCl, pH 7.5–8.0, to a convenient stock (e.g., 10–100 µM). Mix gently; avoid vigorous vortexing that can shear duplexes. Allow 10–15 min for complete dissolution on ice.
• Aliquoting: prepare single‑use aliquots to avoid repeated freeze–thaw. Label with concentration, buffer, and date.
• Storage stability (general):
  • Lyophilized: stable for months at −20°C in a desiccated, dark environment.
  • In solution: store at −20°C (short‑ to medium‑term) or −80°C (long‑term). Avoid more than 3–5 freeze–thaw cycles.
• Handling: maintain RNase‑free workflow; use low‑binding tubes and tips.

Any item‑specific stabilizers, concentration, format, or expiry: Not specified for this item; refer to CoA/Spec Sheet.

Structure and Identity

Item-specific facts (from Product Data):

• Product: AHI1 Human Pre-designed siRNA Set A (SKU: A1487082)
• Intended target: Human AHI1 mRNA (Homo sapiens)
• Storage: Store at -20°C; shipped in ice chest + ice pads
• Molecular formula, exact molecular weight, SMILES, InChIKey: Not specified for this item; refer to CoA/Spec Sheet.

General description (siRNA; literature/general knowledge):

• Short interfering RNA (siRNA) are typically 19–27 bp double‑stranded RNA duplexes with 2‑nt 3′ overhangs per strand, 5′-phosphate and 3′-hydroxyl termini. Many commercial siRNAs are 21‑mer duplexes (sense/antisense) designed to engage the RNA‑induced silencing complex (RISC).
• Structural features: ribose sugars (2′‑OH), phosphodiester backbone (often with partial phosphorothioate or 2′‑O‑Me/2′‑F modifications in some products; not specified for this item), Watson–Crick base pairing forming A‑form helix.
• Set “A” nomenclature commonly denotes one or more distinct siRNA duplexes against non‑overlapping AHI1 sites; the exact number and sequences are not specified for this item.

How to visualize the 2D structure (general):

• Each strand: a linear polymer of ribonucleotides (A, U, G, C) linked 3′→5′ via phosphodiester bonds; the antisense strand is complementary to an AHI1 mRNA region. Duplex forms an antiparallel A‑form helix with 2‑nt 3′ overhangs (e.g., UU).

Synthetic Utility

Not typically applicable: This product is a functional genomics reagent (siRNA) rather than a small‑molecule building block or synthetic intermediate.

General research utility (literature/general):

• Serves as a chemico‑biological tool to modulate gene expression post‑transcriptionally, enabling mechanism studies, pathway mapping, and target validation in human cell systems.
• Complements CRISPR/Cas and shRNA approaches: siRNA permits transient, titratable knockdown with rapid onset, useful when permanent knockout is lethal or triggers compensation.
• Can be integrated into chemical biology workflows: pair with probe compounds, inhibitors, or degraders to dissect on‑target pharmacology vs off‑target effects by observing phenotype rescue or enhancement upon AHI1 silencing.

If your project requires chemical conjugation or custom modifications (e.g., dyes, biotin, or delivery ligands), request custom synthesis; such attributes are not specified for this item.

Target Specificity

Item-specific information:

• Target: AHI1 (Abelson helper integration site 1) mRNA
• Species reactivity: Human (Homo sapiens)
• Product format: Pre‑designed siRNA Set A
• Sequence details, number of duplexes, target exons/coordinates: Not specified for this item; refer to CoA/Spec Sheet.

General notes (literature/general):

• Pre‑designed siRNAs are typically selected to minimize off‑targeting by avoiding seed matches in 3′UTRs and by filtering against SNPs and low‑complexity regions. Validation strategies include dose–response knockdown curves and phenotypic rescue with siRNA‑resistant constructs.