CALB2 Human Pre-designed siRNA Set A

Cat. No.: C1460099

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Overview

CALB2 Human Pre-designed siRNA Set A contains three designed siRNAs for CALB2 gene (Human), as well as a negative control, a positive control, and a FAM-labeled negative control. Components CALB2 siRNA-1: 5 nmol (HPLC) CALB2 siRNA-2: 5 nmol (HPLC) CALB2 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

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

This product requires cold chain shipping. Ground and other economy services are not available.

Product Line

Biological Products

Application Protocols

Item-specific validated applications and dilutions: Not specified for this item; refer to CoA/Spec Sheet.

General RNAi protocol (literature; not product specifications)

1. Preparation

• Thaw siRNA on ice. Briefly spin and keep cold. Work RNase-free.
• Reconstitute to 10–100 µM in RNase-free water or low-EDTA TE. Prepare single-use aliquots and store at −20°C.

2. Transfection complex formation

• Dilute siRNA to the intended working concentration in serum-free medium (e.g., Opti-MEM) per transfection reagent instructions.
• Separately dilute the transfection reagent. Combine and incubate 5–20 min at room temperature to allow complexation.

3. Cell exposure

• Add complexes dropwise to cells at 50–70% confluence to achieve 1–50 nM final siRNA. Swirl gently to distribute.
• Incubate 4–24 h, then replace with complete growth medium.

4. Assessment

• mRNA knockdown: Extract RNA at 24–48 h; perform qRT-PCR using validated primers for CALB2.
• Protein reduction: Harvest at 48–96 h; analyze by Western blot or immunocytochemistry with anti-calretinin antibodies.

5. Controls and data quality

• Include non-targeting control, positive knockdown control, and mock control. Run at least three biological replicates.

6. Optional enhancements

• Reverse transfection for high-throughput formats; electroporation for primary cells with optimized pulse parameters.

Always adapt conditions to your specific cell type and transfection chemistry.

Biological Roles

Focus: CALB2 gene (calbindin 2), also known as calretinin.

General biology (literature; not product specifications)

• Protein class: EF-hand calcium-binding protein of the calbindin family, predominantly expressed in subsets of neurons and certain interneuron populations.
• Cellular roles: Acts as a fast Ca2+ buffer and modulator of intracellular calcium dynamics, influencing neuronal excitability, synaptic integration, and timing-dependent signaling.
• Localization: Mainly cytosolic with potential association to specific subcellular domains where Ca2+ microdomains form.
• Interactions: Binds Ca2+ via EF-hand motifs; may interact with calcium-dependent enzymes and signaling complexes, affecting downstream pathways.
• Functional consequences: Altered CALB2 levels can change calcium waveforms, potentially impacting neurotransmitter release probability and plasticity-related signaling cascades.

Relevance to RNAi experiments

• Knockdown expectations: Reduced CALB2 typically diminishes cytosolic Ca2+ buffering capacity, which can increase peak Ca2+ transients upon stimulation. Phenotypic outcomes are context-dependent and should be validated with appropriate assays (e.g., calcium imaging, electrophysiology).
• Compensation: Cells may exhibit compensatory expression of other calcium-binding proteins; time-course studies and transcript profiling can clarify network adaptations.

Notes

• The above represents general biology to help design experiments with this siRNA set. It does not imply or guarantee any specific outcome in a given model system.

Buffer Applications

Applicability

• Highly relevant: siRNA handling relies on nuclease-free aqueous buffers to preserve integrity and support efficient delivery.

Item-specific

• Supplied buffer and recommended reconstitution medium: Not specified for this item; refer to CoA/Spec Sheet.

General buffer guidance (literature; not product specifications)

• Stock preparation: Reconstitute lyophilized siRNA in RNase-free water or 1× TE (10 mM Tris-HCl, 0.1–1 mM EDTA, pH 7.5–8.0) to 10–100 µM; aliquot and freeze at −20°C.
• Working solutions: Dilute into serum-free, antibiotic-free transfection medium immediately prior to complex formation with the chosen transfection reagent. After 4–24 h, replace with complete growth medium per reagent instructions.
• Salt and pH: Maintain near-physiological pH (7.2–7.6). Moderate monovalent salt (e.g., 100–150 mM Na+) supports duplex stability; avoid high divalent cations before complexation.
• RNase control: Use certified RNase-free buffers and low-binding consumables. Consider adding RNase inhibitors only if compatible with the transfection method.

Example buffer recipes (general)

• 1× TE, low EDTA: 10 mM Tris-HCl, 0.1 mM EDTA, pH 7.5–8.0.
• Nuclease-free PBS for downstream washes (not for initial complexation if it interferes with transfection chemistry).

Quality check

• Measure A260 to confirm concentration post-reconstitution; assess integrity by non-denaturing PAGE if needed.

Green Alternatives

Applicability

• As an aqueous oligonucleotide reagent, the principal environmental considerations relate to buffer selection, plastic consumables, and waste minimization rather than solvent replacement.

General guidance (not product specifications)

• Aqueous systems: Favor RNase-free water or low-EDTA TE for stock solutions. Avoid unnecessary use of organic solvents during associated workflows.
• Process scale: Plan experiments to minimize surplus dilutions, thereby reducing single-use plastics and cold-chain burden.
• Decontamination: Use biodegradable RNase-deactivation solutions where validated; avoid chlorine-based oxidants unless required by facility SOPs.
• Energy use: Consolidate shipping where possible and store at −20°C rather than −80°C when permitted by the product CoA.

Trade-offs

• TE buffer can extend stability (reducing re-synthesis/waste) but EDTA may complicate some delivery systems; balance stability benefits against performance in your transfection method.

Waste handling

• Oligonucleotide solutions are typically non-hazardous; dispose according to institutional guidelines. Segregate sharps and contaminated plastics. Consult the SDS for any product-specific environmental hazards (not specified for this item).

Pharmaceutical Uses

This product is designated For research use only.

Applicability

• Not intended for human or veterinary use, diagnostic procedures, or clinical applications.

General formulation context (literature; not product specifications)

• In pharmaceutical research, siRNAs may be explored as modalities requiring delivery systems (lipid nanoparticles, conjugates). However, such applications demand GMP-grade materials, comprehensive impurity profiling, and rigorous stability/sterility testing—none of which are specified for this item.

Practical implications

• Use this product exclusively for non-clinical laboratory experiments such as target validation, pathway interrogation, and assay development. For any translational or preclinical formulation work, consult suppliers for GMP or “clinic-ready” quality with defined specifications and regulatory documentation.

Physical Properties

Item-specific

• Appearance/physical state: Not specified for this item; refer to CoA/Spec Sheet.
• Concentration, amount, or OD units: Not specified for this item; refer to CoA/Spec Sheet.
• Extinction coefficient (per sequence): Not specified for this item; refer to CoA/Spec Sheet.
• pH of supplied solution (if any): Not specified for this item; refer to CoA/Spec Sheet.

Literature/general (for siRNA; not product specifications)

• Typical length: 19–21 base pairs (∼13–14 kDa per duplex, sequence-dependent).
• Solubility: Freely soluble in RNase-free water or low-salt TE buffer; insoluble in most organic solvents.
• Thermal behavior: siRNA duplexes denature upon heating; melting temperature (Tm) commonly 55–85°C depending on GC content, length, salt, and chemical modifications.
• Ionic effects: Duplex stability increases with monovalent cations (e.g., 50–150 mM Na+ or K+). Mg2+ can further stabilize but may promote aggregation at high concentrations.
• Storage form: Often provided lyophilized or in aqueous, nuclease-free buffer; stable at −20°C when protected from RNases and repeated freeze–thaw.

Practical notes

• Reconstitution is typically in RNase-free water or 1× TE (10 mM Tris, 0.1–1 mM EDTA, pH 7.5–8.0) to 10–100 µM stock; exact instructions are Not specified for this item; refer to CoA/Spec Sheet.
• Avoiding divalent-cation-rich buffers and maintaining low EDTA helps compatibility with cationic transfection reagents.

Quality & Grades

Item-specific

• Grade/purity: Not specified for this item; refer to CoA/Spec Sheet.
• Stabilizers or modifications (e.g., 2′-OMe, PS): Not specified for this item; refer to CoA/Spec Sheet.

Context and expectations for siRNA quality (general; not product specifications)

• Synthesis quality: Research-grade siRNAs are typically solid-phase synthesized and deprotected; optional cartridge or HPLC purification reduces truncated sequences. HPLC-grade siRNA exhibits lower levels of n − 1 species and improved performance in sensitive applications.
• Identity confirmation: Mass spectrometry (MALDI-TOF/ESI) and analytical HPLC/UPLC are commonly used. OD260 measurement verifies quantity but not integrity.
• Functional quality: Potency depends on sequence design, off-target minimization, and duplex thermodynamics. Pre-designed sets often include multiple non-overlapping duplexes to increase the chance of robust knockdown.
• RNase-free assurance: Packaging and QC in nuclease-free environments are critical. Endotoxin specifications may be relevant for primary cell work (not specified for this item).

Implications

• If grade/purity are critical (e.g., for challenging cell types), consult the CoA/Spec Sheet for purification level, residual salts/solvents, and analytical traces. Absent item-specific specifications, users should perform a small-scale pilot to evaluate knockdown efficacy and cytotoxicity with their transfection system.

Reaction & Applications

This product is used for RNA interference (RNAi)-mediated transient knockdown of human CALB2 transcripts in cell-based or cell-free systems.

Item-specific

• Target: Human CALB2 (calbindin 2, calretinin). Additional target details (e.g., transcript IDs, exact target sites) are Not specified for this item; refer to CoA/Spec Sheet.

General applications (literature; not product specifications)

• Gene function analysis: Evaluate CALB2 roles in calcium buffering, neuronal excitability, or signaling pathways by quantifying mRNA/protein reduction after transfection.
• Pathway dissection: Combine with agonists/antagonists or calcium imaging to assess downstream effects of CALB2 depletion.
• Assay modalities: qRT-PCR for mRNA knockdown, Western blot or immunocytochemistry for protein, and phenotypic readouts (e.g., electrophysiology, Ca2+ imaging) depending on model.
• Controls: Non-targeting siRNA; positive control siRNA against a housekeeping or reporter gene; mock transfection for baseline cytotoxicity.

Practical notes

• Delivery: Lipid- or polymer-based transfection in adherent lines; electroporation for suspension or primary cells. Optimize siRNA dose (commonly 1–50 nM final) and reagent ratios empirically.
• Timing: Peak mRNA knockdown often at 24–48 h post-transfection; protein knockdown may lag based on protein half-life.
• Replicates and statistics: Use biological triplicates and include transfection efficiency markers where feasible.

Limitations

• Off-target effects and innate immune activation are mitigated by careful dosing, appropriate controls, and validated readouts.

Reaction Conditions

Interpretation for siRNA experiments

• Here, “reaction conditions” refer to transfection and gene-silencing conditions rather than chemical reaction parameters.

Item-specific

• Recommended cell lines, transfection reagent, dose, or exposure time: Not specified for this item; refer to CoA/Spec Sheet.

General conditions (literature; not product specifications)

• Cell density: Plate adherent cells to 50–70% confluence at transfection to balance uptake and viability.
• Dosing: Typical final siRNA concentrations range 1–50 nM, optimized per cell type and delivery method.
• Complexation: Mix siRNA with lipid/polymer transfection reagent in serum-free medium; incubate 5–20 min to form complexes before adding to cells.
• Exposure: 4–24 h in transfection medium, then replace with complete medium; assess mRNA at 24–48 h and protein at 48–96 h or per protein half-life.
• Controls: Include non-targeting siRNA, positive knockdown control, and mock (reagent-only) control.
• Readouts: qRT-PCR (ΔΔCt) for mRNA; Western blot or ICC/IF for protein; functional assays tailored to CALB2 biology.

Troubleshooting

• Low knockdown: Titrate dose, switch transfection reagent, verify cell health, and confirm sequence identity.
• Cytotoxicity: Reduce reagent amount, shorten exposure, or use gentler delivery methods (e.g., electroporation parameters optimized to maintain viability).

Safety & Handling

Authoritative safety information must be taken from the product SDS. The following summarizes general best practices for handling synthetic siRNA reagents in the laboratory.

Item-specific (from Product Data)

• GHS classification, pictograms, signal word, H-statements: Not specified for this item; refer to SDS.
• Storage conditions: Store at −20°C.
• Shipping: Shipped in ice chest with ice pads.
• Intended use: For research use only.

General laboratory safety for oligonucleotides (not product specifications)

• PPE: Lab coat, gloves, and eye protection. Change gloves frequently to avoid RNase contamination.
• RNase control: Use certified RNase-free tubes, tips, and solutions. Work in clean areas; decontaminate benches and instruments with RNase-deactivating solutions.
• Incompatibilities: Avoid strong acids/bases and nucleases; minimize exposure to elevated temperatures and repeated freeze–thaw cycles. Keep solutions salt-balanced to prevent aggregation/precipitation.
• First aid (general): If contact with eyes/skin occurs, rinse with water. If inhaled (powder), move to fresh air. If ingested, rinse mouth with water. Seek medical attention as needed. Always follow SDS guidance.
• Waste: Dispose of according to institutional biosafety and chemical waste procedures for non-hazardous biological reagents unless otherwise specified by SDS.

Special risks

• Microbial contamination and RNase introduction are the principal risks to integrity and performance. Maintain aseptic, nuclease-free technique throughout handling and experiment setup.

Solvent Selection

Applicability

• As an oligonucleotide duplex, this product is handled in aqueous, RNase-free media rather than organic solvents.

Item-specific

• Recommended solvent/buffer for reconstitution: Not specified for this item; refer to CoA/Spec Sheet.

Literature/general guidance (not product specifications)

• Primary choices: RNase-free water for short-term use; 1× TE (10 mM Tris, 0.1–1 mM EDTA, pH 7.5–8.0) for longer-term stability by chelating trace divalent cations.
• Working dilutions: Prepare in serum-free, antibiotic-free transfection medium immediately prior to complexation with lipid/polymer reagents; switch to complete medium after 4–24 h, per transfection reagent guidance.
• Avoid: High concentrations of divalent cations (Mg2+, Ca2+) prior to complexation; organic solvents; buffers containing RNases.

Comparison (general)

• Water vs TE: Water maximizes compatibility with all transfection reagents but offers less protection from hydrolysis/metal-catalyzed degradation. TE provides improved stability for frozen stocks but EDTA may interfere with some cationic delivery systems at high concentrations—use low-EDTA TE.

Practical tips

• Filter-sterilize buffers and confirm RNase-free status. Thaw on ice; mix gently to avoid shearing or foaming. Use low-binding plastics to minimize adsorption losses at low nM dilutions.

Storage & Reconstitution

Item-specific (from Product Data)

• Storage conditions: Store at −20°C.
• Shipping: Ice chest with ice pads.
• Supplied form, concentration, stabilizers, and reconstitution instructions: Not specified for this item; refer to CoA/Spec Sheet.

General guidance for siRNA (literature; not product specifications)

• Upon receipt: If lyophilized, briefly centrifuge before opening to collect material at the bottom of the vial.
• Reconstitution: Dissolve in RNase-free water or 1× TE (10 mM Tris, 0.1–1 mM EDTA, pH 7.5–8.0) to 10–100 µM. Mix gently; avoid vigorous vortexing that can introduce foaming.
• Aliquoting: Prepare small, single-use aliquots to prevent repeated freeze–thaw. Use low-binding tubes to minimize adsorption.
• Storage: Keep aliquots at −20°C protected from light and moisture. For long-term storage, −80°C may further preserve integrity if permitted by the CoA.
• Freeze–thaw: Minimize cycles; thaw on ice and keep cold during setup.
• Stability checks: Verify concentration by A260; inspect for precipitation or turbidity. If performance declines, prepare fresh working dilutions from frozen stock.

Compatibility notes

• Avoid high-EDTA stocks if using cationic delivery systems sensitive to chelation. Ensure buffers are RNase-free. For cell transfection, prepare working dilutions immediately before complexation.

Structure & Identity

Overview: CALB2 Human Pre-designed siRNA Set A consists of double-stranded small interfering RNAs intended to transiently silence the human CALB2 gene (calbindin 2, calretinin) in research settings.

Item-specific (from Product Data)

• Product type: Pre-designed siRNA set targeting human CALB2
• Sequence information: Not specified for this item; refer to CoA/Spec Sheet.
• Number of duplexes in “Set A”: 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.

Literature/general characteristics of siRNA (not product specifications)

• Typical architecture: 19–21 bp RNA duplex with 2-nt 3′ overhangs on each strand; antisense (guide) strand complementary to target mRNA; sense (passenger) strand matching the mRNA.
• Common sugar/base chemistry: 2′-OH ribose; uracil instead of thymine; optional phosphorothioate or 2′-O-modifications in some products (not specified for this item).
• Representative descriptors: SMILES, InChIKey, and exact composition are sequence-dependent and therefore not universally defined for siRNA mixtures.

2D structural description (general)

• Two antiparallel RNA strands aligned in A-form helical geometry, each nucleotide containing a phosphate backbone, ribose sugar, and a heteroaromatic nucleobase (A, U, G, C). The duplex includes short 3′ overhangs that aid RISC loading and enhance silencing efficiency.

Notes

• Any identifiers such as CAS, CID, SMILES, InChIKey, and exact MW are sequence-dependent and are Not specified for this item; refer to CoA/Spec Sheet.

Synthetic Utility

Not typically applicable. As a biological reagent (siRNA), this product is not used as a synthetic building block or reagent in organic synthesis. For experimental considerations relevant to this product type, see Reaction & Applications, Buffer Applications, and Application Protocols.

Target Specificity

Item-specific (from Product Data)

• Target name: CALB2 Human (calbindin 2; calretinin)
• Product type: Pre-designed siRNA Set A
• Species reactivity: Human (implied by product name)
• Sequence details (guide/sense strands), target exons/transcripts, and number of duplexes: Not specified for this item; refer to CoA/Spec Sheet.

Notes

• Off-target minimization, strand bias optimization, and seed-region design are critical to specificity but are not described for this particular item. Users should validate knockdown by qRT-PCR and protein assays and include appropriate non-targeting and positive controls.

Frequently Asked Questions

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.