Recombinant EIF3E Antibody - Primary antibody, specific to EIF3E

Cat. No.: Ab327186

GRADE & PURITY KD Validation

Application

Flow Cytometry, WB

Host species

Rabbit

Reactive species

Human, Mouse, Rat

Isotype

Rabbit IgG

Conjugation

Unconjugated

Specifications

Product Name

Recombinant EIF3E Antibody - Primary antibody, specific to EIF3E

Specifications & Purity

KD Validation

Host species

Rabbit

Specificity

EIF3E

Immunogen

A synthesized peptide derived from human eIF3e

Conjugation

Unconjugated

Grade

KD Validation

Purification Method

Protein A purified

Storage and Shipping

Storage

Store at -20°C,Avoid repeated freezing and thawing

Shipped In

Ice chest + Ice pads

Stability And Storage

Store at -20°C for one year.

Application Protocols

• Item-specific (Product Data):

  • Tested applications, recommended dilutions, positive control lysates/tissues, and antigen retrieval conditions: Not specified for this item; refer to CoA/Spec Sheet.

• General protocols (literature; to aid initial setup only):

  • Western blot (WB): Block PVDF membrane with 5% BSA in TBS-T for 1 h; incubate with primary anti‑EIF3E antibody (optimize starting range e.g., 1:500–1:2,000) overnight at 4°C; wash and incubate with HRP-conjugated secondary (1:5,000–1:20,000) for 1 h; develop via ECL.
  • Immunoprecipitation (IP): Incubate clarified lysate (0.5–2 mg total protein) with antibody for 2 h to overnight at 4°C; add Protein A/G beads for 1 h; wash 3–5×; elute with SDS sample buffer.
  • Immunofluorescence (IF/ICC): Fix cells, permeabilize with 0.1–0.3% Triton X-100, block with 1–5% BSA, incubate primary 1–2 h RT or overnight 4°C (starting 1:200–1:1,000), secondary 1 h with appropriate fluorophore; mount with anti-fade medium.

These conditions are starting points and require optimization. Use of this SKU in any specific application should be guided by the product’s datasheet.

Biological Roles

Context on the target (literature; not a property of the antibody itself):

• EIF3E (aka INT6) is a subunit of the eukaryotic translation initiation factor 3 (eIF3) complex, which facilitates recruitment of the 40S ribosomal subunit to mRNA and stabilizes the preinitiation complex.
• Functional notes (literature):
  • EIF3E contributes to assembly/stability of the eIF3 complex and interacts with other subunits to regulate translation initiation efficiency.
  • It has been implicated in coupling translation with protein quality control and in cellular stress responses through modulation of translation programs.
  • Subcellular localization is predominantly cytosolic, with association to ribosomes and translation initiation assemblies; context-dependent nuclear roles have been reported.
• Orthology: EIF3E is conserved across eukaryotes; cross-reactivity of antibodies may vary by sequence conservation and should be empirically verified.

How this relates to antibody use (general):

• Antibodies against EIF3E are employed to quantify expression changes, monitor pathway modulation in translation studies, and isolate eIF3-containing complexes.
• KD validation, when available, strengthens confidence that observed signals derive from EIF3E.

Note: No clone, epitope, or species reactivity is specified for this item; consult the product documentation for target sequence coverage and tested species.

Buffer Applications

This product is an antibody and is not a buffering reagent. Therefore, classical buffer system formulation (pKa selection, capacity) is not applicable.

• Item-specific (Product Data):

  • Supplied buffer composition: Not specified for this item; refer to CoA/Spec Sheet.

• Practical notes for users (general guidance):

  • Working buffers: For immunoassays, PBS or TBS at pH ~7.2–7.6 with 0.01–0.05% Tween-20 is commonly used for washes; blocking buffers may include 3–5% BSA or nonfat dry milk depending on assay and detection enzyme.
  • Sample prep: Maintain compatible salt and detergent levels to preserve antigen–antibody interactions; avoid chelators or high detergent levels that disrupt binding unless assay-specific.

For precise formulations and compatibility with this SKU, consult the CoA/Spec Sheet.

Green Alternatives

Antibody reagents have a modest direct environmental impact compared to bulk solvents, but sustainability can be improved through formulation, handling, and logistics.

• Item-specific (Product Data):

  • Shipped in ice chest + ice pads. Storage at –20°C.

• Greener practice options (general; not SKU-specific):

  • Shipping/logistics: Coordinate consolidated shipments and choose ground shipping when timelines allow to reduce carbon footprint. Reuse/return insulated shippers where permitted.
  • Formulation: Preservative-free or low-sodium buffers may reduce downstream waste for sensitive applications; verify stability trade-offs. High-concentration stocks reduce packaging and cold-chain volume.
  • Waste minimization: Aliquot appropriately to avoid discarding thawed excess; adopt validated micro-volume protocols to reduce reagent consumption.
  • Energy efficiency: Store at the warmest validated condition (–20°C vs –80°C if stability allows) and periodically defrost freezers for efficiency.
  • Alternative detection: Use highly sensitive detection chemistries (e.g., enhanced chemiluminescence or near-IR fluorophores) to lower primary antibody usage per assay.

• Trade-offs:

  • Preservative-free formats may require stricter aseptic technique and shorter open-vial lifetimes.
  • Reduced packaging must not compromise temperature control; ensure cold-chain integrity to prevent performance loss.

These measures complement, but do not replace, product-specific guidance in the CoA/SDS.

Pharmaceutical Uses

This product is supplied strictly for research use only (Product Data). It is not intended for diagnostic, therapeutic, or clinical applications.

• Item-specific (Product Data):

  • Research Use Note: For research use only.

• General information (non-clinical):

  • Antibodies can serve as analytical reagents in bioprocess development (e.g., ELISAs for process monitoring) and as reference tools in characterization of translation initiation pathways in discovery research.
  • No pharmacopeial monograph, excipient status, or GMP suitability is provided for this item.

• Compliance note:

  • Users are responsible for ensuring the reagent is appropriate for their laboratory’s intended research use and regulatory environment. Do not incorporate into products for human or veterinary use.

Physical Properties

Summary: As a protein reagent, conventional small‑molecule physico-chemical constants (bp, mp, logP) are not applicable.

• Item-specific (Product Data):

  • Appearance: Not specified for this item; refer to CoA/Spec Sheet.
  • Concentration, buffer composition, preservative content: Not specified for this item; refer to CoA/Spec Sheet.

• Non-applicable parameters for antibodies:

  • Boiling point, melting point, vapor pressure, logP, SMILES-based descriptors: Not applicable to macromolecular protein solutions.

• General literature values and considerations (not specific to this SKU):

  • Apparent molecular mass (IgG): ~150 kDa by SDS-PAGE under non-reducing conditions; heavy chain ~50 kDa and light chain ~25 kDa under reducing conditions.
  • Solubility: Antibodies are soluble in aqueous buffers (e.g., PBS, TBS) across pH ~6.5–8.0; stability depends on ionic strength, pH, and excipients (e.g., 0.01–0.1% Tween-20 to reduce adsorption; 0.1–5% BSA/trehalose as carriers).
  • Isoelectric point (pI): IgG molecules typically span pI ~6–9 depending on sequence and glycosylation.
  • Temperature sensitivity: Activity diminishes with repeated freeze–thaw and prolonged exposure to >4–8°C (literature); hence, storage at –20°C or below in aliquots is common.
  • Light sensitivity: Extended UV exposure can induce oxidation and aggregation; protect from light when feasible.

• Practical note: For any quantitative handling (e.g., molar dosing), obtain the exact concentration and format from the product’s CoA/Spec Sheet.

Quality & Grades

• Item-specific (Product Data):

  • Grade/Purity: KD Validation.

• What “KD Validation” typically means (general explanation; not a guarantee for unlisted applications):

  • KD (knockdown) validation refers to demonstrating antibody specificity by reducing the target’s expression (e.g., siRNA/shRNA/CRISPRi against EIF3E) and showing a corresponding loss or reduction of the detected signal (e.g., in Western blot or IF). This provides orthogonal evidence that the observed band/signal arises from EIF3E rather than off-target proteins.
  • Advantages: Enhances confidence in target specificity, reduces false positives, and supports reproducible data.

• Additional quality dimensions (not specified for this item; refer to CoA/Spec Sheet):

  • Isotype, clonality (mono/polyclonal), host species, recombinant format (full-length IgG vs fragment), affinity/avidity metrics (KD by SPR/BLI), endotoxin level, purity by SDS-PAGE/HPLC, residual host cell proteins/DNA, and preservative content.

• Practical implications for users:

  • When planning experiments beyond the validation context (e.g., IP, IHC, FC), verify application suitability, working dilutions, and species reactivity from the product’s documentation.
  • For quantitative work, confirm lot-specific concentration and recommended loading/antigen retrieval conditions where relevant.
  • Maintain consistent handling and storage to preserve the validated performance characteristics.

For authoritative details on this SKU’s QC attributes, consult the CoA/Spec Sheet.

Reaction & Applications

As a biological antibody reagent, this product is not used as a reagent in synthetic chemical reactions. Its relevance lies in immunoassay and protein analysis workflows.

• Item-specific (Product Data):

  • Manufacturer Applications: Not specified in the listing; see general applications below as literature guidance only.

• General research applications for EIF3E antibodies (literature; not specific to this SKU):

  • Western blot (WB): Detect EIF3E in cell/tissue lysates to assess expression levels; KD validation typically shows signal reduction upon EIF3E knockdown.
  • Immunoprecipitation (IP)/Co-IP: Enrich EIF3E and associated eIF3 complex members to study protein–protein interactions and translation-initiation assemblies.
  • Immunofluorescence (IF)/Immunocytochemistry (ICC): Visualize subcellular localization of EIF3E in cultured cells; fixation/permeabilization conditions require optimization.
  • Proximity assays: Use in PLA or BioID follow-ups to validate interactions identified by proteomics.
  • ELISA/Sandwich assays: Potentially applicable if paired with a complementary capture/detection antibody; format must be verified for this SKU.

• Practical tips (general):

  • Verify knockdown/knockout controls to confirm specificity; include secondary-only and isotype controls to assess background.
  • Optimize antibody dilution, incubation times, and antigen retrieval/fixation depending on application.
  • Maintain cold chain and avoid freeze–thaw cycles to preserve binding activity.

Reaction Conditions

Classical reaction conditions (solvent, temperature, catalysts) do not apply to antibody reagents. Below are general assay-condition guidelines for immunoapplications (literature; not specific to this SKU):

• Western blot:

  • Sample: Denatured lysates; 20–40 µg total protein per lane typical.
  • Primary incubation: Overnight at 4°C or 1–2 h at room temperature; dilution to be optimized empirically.
  • Wash: TBS-T or PBS-T, 3–5 times, 5–10 min each.

• Immunoprecipitation:

  • Lysis: Non-denaturing buffers (e.g., 20–50 mM Tris, 150 mM NaCl, 0.5–1% NP-40/Triton X-100), protease/phosphatase inhibitors.
  • Incubation: 1–4 h at 4°C with rotation; capture with Protein A/G beads matched to antibody isotype.

• IF/ICC:

  • Fixation: 4% paraformaldehyde or cold methanol (optimize); permeabilization with 0.1–0.3% Triton X-100 if needed.
  • Blocking: 1–5% BSA or serum for 30–60 min; primary incubation 1–2 h RT or overnight 4°C.

• ELISA (sandwich/indirect):

  • Coating: 0.5–2 µg/mL capture Ab overnight at 4°C; detection Ab/secondary per platform.

• Notes:

  • Adjust conditions based on antigen abundance and antibody affinity.
  • Always include appropriate controls: isotype, secondary-only, and KD/KO where available.

Item-specific recommended dilutions or validated applications are not provided; refer to CoA/Spec Sheet.

Safety & Handling

Overview: Protein antibodies generally present low acute toxicity; however, formulations can contain preservatives, stabilizers, or buffer salts that require prudent handling.

• Item-specific (Product Data):

  • GHS Classification, Signal Word, H-Statements, Pictograms: Not specified for this item; refer to SDS.
  • Storage: Store at –20°C; avoid repeated freezing and thawing.
  • Shipment: Ice chest + ice pads.

• General safety guidance (literature/best practice):

  • Personal protective equipment (PPE): Lab coat, nitrile gloves, and eye protection when handling liquids to avoid skin/eye contact.
  • Hygienic measures: Avoid aerosol generation and ingestion; wash hands after use.
  • First aid (general): If contact occurs, rinse affected area with water; if inhaled as aerosol, move to fresh air; seek medical attention as needed. Always consult the SDS for authoritative instructions.
  • Incompatibilities: Avoid strong oxidizers and proteases; do not mix with microbial contaminants. Maintain aseptic technique to prevent degradation.
  • Physical risks: Repeated freeze–thaw cycles can cause aggregation/precipitation; maintain cold chain to preserve activity.
  • Environmental: Dispose of proteinaceous waste in accordance with institutional biosafety procedures; do not release into drains without approval.

• Spill/cleanup:

  • Absorb small spills with inert material; decontaminate surfaces with suitable detergent or 70% ethanol; dispose according to local regulations.

Always defer to the product-specific SDS/CoA for definitive hazard and handling details.

Solvent Selection

This product is an aqueous protein reagent; classical organic solvent selection is generally not applicable.

• Item-specific (Product Data):

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

• General guidance for antibody handling (literature; not specific to this SKU):

  • Preferred media: Phosphate-buffered saline (PBS) or Tris-buffered saline (TBS), pH ~7.2–7.6, often with 0.01–0.05% Tween-20 to minimize nonspecific adsorption during immunoassays.
  • Stabilizers: 0.1–1% BSA, gelatin, or 2–5% trehalose can improve freeze–thaw and long-term stability. Verify compatibility with your downstream assays (e.g., avoid BSA for certain mass-spec workflows).
  • Avoid: High concentrations of organic solvents (acetone, acetonitrile, methanol, DMSO) that can denature proteins; if necessary for specific protocols, keep organic content minimal and validate recovery.
  • Detergents/chaotropes: Small amounts of nonionic detergents (Tween-20, Triton X-100) can aid in blocking and wash steps; strong chaotropes (urea, guanidinium) may disrupt binding and should be avoided unless used for elution in IP.

• Comparison (general):

  • PBS vs TBS: TBS can reduce background in alkaline phosphatase-based detection; PBS is broadly compatible and commonly used in HRP-based systems.

Always check the CoA/Spec Sheet for the supplied buffer and adjust only if validated for your assay.

Storage & Reconstitution

• Item-specific (Product Data):

  • Storage Conditions: Store at –20°C. Avoid repeated freezing and thawing.
  • Shipped In: Ice chest + ice pads (maintain cold chain upon receipt).

• If liquid format (not specified for this item):

  • Upon receipt, briefly centrifuge to collect contents; aliquot into single‑use volumes to prevent freeze–thaw cycles. Store aliquots at –20°C (or as otherwise indicated on CoA/Spec Sheet).

• If lyophilized format (not specified for this item):

  • Reconstitute with sterile, antibody‑grade water or PBS to the desired concentration; gently invert to mix—do not vortex vigorously. Allow to fully dissolve on ice.

• General stability guidance (literature; not specific to this SKU):

  • Short-term storage: 2–8°C for days to weeks may be acceptable if preservative is present; verify on datasheet.
  • Long-term storage: –20°C to –80°C recommended depending on formulation; addition of carrier proteins (e.g., 0.1% BSA) and cryoprotectants (e.g., 10–50% glycerol) can enhance stability, subject to compatibility with assays.
  • Avoid repeated freeze–thaw; minimize exposure to ambient temperatures and light.

• Handling tips:

  • Use low-protein-binding tubes and tips to reduce loss.
  • Maintain aseptic technique to prevent contamination.

For exact buffer composition, concentration, and reconstitution instructions for this SKU, refer to the CoA/Spec Sheet.

Structure & Identity

Brief overview: Recombinant EIF3E Antibody is a protein-based biological reagent targeting the eukaryotic translation initiation factor 3 subunit E (EIF3E).

• Item-specific (Product Data):

  • Product Name: Recombinant EIF3E Antibody (SKU Ab327186)
  • Grade/Purity: KD Validation (see Quality & Grades)
  • 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 / CAS / CID: Not applicable to protein biologics and not specified for this item.

• Structural features (general literature context, not specific to this SKU):

  • Antibodies are Y‑shaped glycoproteins composed of two heavy and two light chains linked by disulfide bonds, forming variable (V) regions that confer antigen specificity and constant (C) regions that define isotype.
  • Recombinant antibodies are produced from defined DNA sequences in mammalian or other expression systems, offering lot-to-lot consistency versus hybridoma-derived materials.
  • Typical full-length IgG has an apparent MW ~150 kDa (literature); fragment formats (Fab, scFv, VHH) are smaller, but the format for this item is not specified.

• Antigen identity (general):

  • EIF3E (also known as INT6) is a subunit of the eIF3 complex involved in translation initiation; cellular localization is predominantly cytosolic with association to ribosomal machinery (literature). No epitope, clone, or species reactivity is specified for this item.

Synthetic Utility

Not applicable to this product type. As a recombinant antibody, it is not employed as a building block or reagent in synthetic organic chemistry.

• For users seeking chemical tools: Consider small-molecule probes targeting translation initiation pathways or tagged peptide antigens for assay calibration.

• Relevant utility (biochemical, general): Antibodies function as affinity reagents to capture, detect, or quantify EIF3E in complex biological matrices, enabling downstream analyses (WB, IP, IF, ELISA).

Target Specificity

• Item-specific (Product Data):

  • Antigen/epitope, clone ID, isotype, host species, and tested species reactivity: Not specified for this item; refer to CoA/Spec Sheet.

• General context (literature; for background only):

  • Intended target: EIF3E (INT6), a component of the eIF3 complex involved in translation initiation.
  • Expected subcellular localization of target: Cytosol, associated with ribosomal 40S subunits and translation preinitiation complexes.
  • Expected molecular weight of target (human EIF3E): ~52 kDa by sequence; apparent migration can vary by isoform/post-translational modifications (literature).

• Specificity considerations (general):

  • KD validation typically shows reduced signal in cells with EIF3E knockdown/knockout, supporting specificity.
  • Sequence homology across species may yield cross-reactivity; verify target peptide alignment if using in non-human models.

For this SKU, consult the product datasheet for definitive specificity, clone/epitope, and species information.