WTAP Mouse mAb - Primary antibody, specific to WTAP

Cat. No.: Ab325644

GRADE & PURITY KD Validation

Application

Flow Cytometry, IF/ICC, WB

Host species

Mouse

Reactive species

Human, Mouse, Rat

Isotype

Mouse IgG1

Conjugation

Unconjugated

Specifications

Product Name

WTAP Mouse mAb - Primary antibody, specific to WTAP

Spécifications et pureté

KD Validation

Espèce hôte

Mouse

Spécificité

WTAP

Immunogène

Recombinant protein of human WTAP

Conjugation

Unconjugated

Grade

KD Validation

Méthode de purification

Protein A/G purified

Product Line

Produits biologiques

Stockage et expédition

Conditions de stockage de stockage

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

Expédié en

Ice chest + Ice pads

Stabilité et stockage

Store at -20°C for one year.

Application Protocols

• Item-specific tested applications and recommended dilutions

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

• Positive/negative controls

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

• Secondary antibodies and detection systems

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

• Support

  • For protocol recommendations tailored to your platform (WB, IP, IF, IHC, etc.), please contact technical support with your species, sample type, and detection method.

Biological Roles

• Target overview (general literature on WTAP; not item-specific claims)

  • WTAP (Wilms Tumor 1‑Associating Protein) is a nuclear protein that localizes to nuclear speckles and is integral to the m6A RNA methyltransferase (writer) complex, partnering with METTL3/METTL14 and other adaptors. WTAP contributes to RNA N6‑methyladenosine (m6A) deposition, impacting mRNA processing, stability, and translation.
  • Beyond m6A regulation, WTAP has reported roles in pre‑mRNA splicing, cell cycle control, and developmentally regulated gene expression (literature).

• Utility of WTAP detection (general research rationale)

  • Monitoring WTAP levels and localization aids studies of epitranscriptomic regulation and RNA metabolism.
  • Perturbation experiments (e.g., siRNA/CRISPR) combined with immunoassays enable pathway dissection and validation of mechanistic hypotheses.

• Cautions

  • WTAP exists in multi‑protein complexes; co‑migration or co‑enrichment of partners can occur in IP/RIP experiments—use proper controls and orthogonal readouts.

• Item-specific statement

  • This product’s name indicates specificity to WTAP, but detailed epitope, cross‑reactivity, and validated applications are not specified for this item; refer to CoA/Spec Sheet.

Buffer Applications

• Item-specific

  • Exact storage or assay buffer composition is not specified for this item; refer to CoA/Spec Sheet.

• General antibody buffer guidance (literature; not item-specific)

  • Storage buffers: PBS or TBS at pH 7.2–7.6; optional 10–50% glycerol to protect from freeze–thaw; optional carrier protein (e.g., 0.1% BSA) to reduce adsorption if compatible with assays.
  • Assay buffers:
    • WB: Tris‑glycine or Bis‑Tris running buffers; TBST (TBS + 0.1% Tween‑20) for washes; 5% nonfat milk or 3% BSA for blocking.
    • IF/ICC: PBS or HBSS for washes; blocking with 1–5% BSA/serum; permeabilization with 0.1–0.3% Triton X‑100 or saponin depending on target localization.
    • IP: Lysis buffers such as NP‑40 or RIPA variants with protease/RNase inhibitors; adjust stringency to preserve relevant complexes.
  • pH considerations: Maintain near‑neutral pH to preserve antibody structure and antigen integrity; avoid chelators if metal‑dependent interactions are assayed.

• Practical tips

  • Filter‑sterilize and use low‑bind tubes to minimize protein loss.
  • Validate detergent type and concentration for compatibility with downstream detection (e.g., mass spectrometry or RNA‑binding assays).

Green Alternatives

• Context

  • Antibodies are aqueous biological reagents; “green” considerations focus on formulation, cold‑chain logistics, and waste minimization rather than solvent substitution.

• General greener practices (literature/general guidance)

  • Formulation optimization: Prefer azide‑free buffers when possible to reduce hazardous waste, especially for flow cytometry or live‑cell contexts. If antimicrobial protection is needed, consider low‑toxicity alternatives validated for your assay.
  • Lyophilized formats: When available, can reduce shipping weight/volume and reliance on cold packs; reconstitute with sterile buffer at use site.
  • Concentrate and aliquot: Smaller aliquots reduce freeze–thaw losses and product waste.
  • Cold‑chain efficiency: Utilize phase‑change materials and consolidated shipments to lower carbon footprint while maintaining integrity.
  • Waste handling: Segregate proteinaceous and azide‑containing waste; follow institutional green-chemistry disposal routes where available.

• Comparative considerations (example; not item-specific)

  • Liquid with preservative vs Lyophilized, preservative‑free:
    • Liquid: ready‑to‑use; requires continuous refrigeration/freezing; potential hazardous preservative.
    • Lyophilized: reduced shipping burden and no preservative; requires careful reconstitution and aliquoting.

• Item-specific note

  • This item ships with an ice chest + ice pads and is stored at −20°C (from Product Data). Alternative formats are not specified for this item.

Pharmaceutical Uses

• Applicability note

  • This product is labeled For research use only (from Product Data). It is not intended for diagnostic, therapeutic, or clinical applications.

• General roles of antibodies in pharmaceutical R&D (literature; not item-specific)

  • As analytical reagents in process development and quality control (e.g., host‑cell protein assays, target expression monitoring) when appropriately qualified.
  • In mechanism‑of‑action studies and biomarker discovery workflows to characterize targets and pathways.
  • For immunocapture in LC–MS sample prep to enrich low‑abundance proteins.

• Compliance considerations

  • Transition from research to regulated use requires extensive qualification/validation (specificity, linearity, precision, robustness) and documentation (SOPs, change control). Those activities are beyond the scope of this catalog listing.

• Item-specific statement

  • No pharmacopeial status, excipient role, or GMP manufacturing information is provided for this item.

Physical Properties

• Item-specific (from Product Data)

  • Appearance: Not specified for this item; refer to CoA/Spec Sheet.
  • Concentration / Formulation / Buffer salts / Preservatives: Not specified for this item; refer to CoA/Spec Sheet.
  • Molecular Weight / Formula: Not specified for this item; refer to CoA/Spec Sheet.

• General/literature properties of monoclonal antibodies (for laboratory handling; not item-specific)

  • Macromolecular class: Protein (immunoglobulin). Typical IgG MW ~150 kDa (literature).
  • Solubility: Readily soluble in aqueous buffers such as PBS or TBS; incompatible with strong denaturants unless intended for SDS‑PAGE.
  • Stability: Sensitive to repeated freeze–thaw; stability typically enhanced by glycerol (e.g., 10–50%) and protein stabilizers (e.g., BSA) when appropriate for the application.
  • pI (isotype/clone dependent): Often in the ~6–9 range (literature), affecting binding to ion‑exchange media.
  • Optical properties: Protein UV absorbance maximum at 280 nm due to aromatic residues; A280 can be used to estimate concentration (literature).
  • Storage temperature dependence: Long‑term storage typically at −20 to −80°C (formulation dependent); short‑term at 2–8°C (literature guidance).

• Practical implications

  • Avoid surface adsorption and aggregation by using low‑bind plastics and gentle mixing.
  • Maintain neutral pH and isotonicity to reduce denaturation. Exact values for this specific item are not provided.

Quality and Grades

• Item-specific (from Product Data)

  • Grade/Purity: KD Validation.

• Interpretation of KD Validation (general meaning; not a regulatory grade)

  • KD (knockdown) validation typically indicates that the antibody’s on‑target specificity has been corroborated by reducing or eliminating target expression (e.g., via siRNA/shRNA or CRISPR) and observing a corresponding loss or diminution of the antibody signal. This provides strong orthogonal evidence that the detected band or staining arises from the intended antigen.
  • KD validation complements other assessments (e.g., peptide competition, isotype controls, recombinant protein controls) but does not, by itself, define buffer composition, purity, or application suitability.

• Additional quality notes (applicable considerations)

  • For antibodies, quality encompasses: correct banding pattern in Western blot, expected subcellular localization in IF/IHC, absence of off‑target bands at validated loads, and lot‑to‑lot consistency.
  • If specific metrics (e.g., host species, isotype, clonality/clone ID, concentration, preservative levels, endotoxin) are critical to your workflow, please refer to the CoA/Spec Sheet or contact technical support for lot‑resolved data.

• Documentation

  • This item is labeled For research use only (from Product Data). No clinical or diagnostic use is implied.

Reaction and Applications

• Note on applicability

  • Classical organic reaction chemistry is not applicable to antibody reagents. Instead, applications are immunochemical.

• General research applications for a WTAP-targeting mouse monoclonal antibody (literature/general guidance; not item-specific validation)

  • Western blot (WB): Detection of WTAP in cell/tissue lysates under denaturing conditions; expect a band consistent with WTAP’s predicted size when samples express the protein.
  • Immunoprecipitation (IP): Enrichment of WTAP and associated complexes (e.g., components of the m6A writer complex) for downstream proteomics or activity assays.
  • Immunofluorescence/Immunocytochemistry (IF/ICC): Visualization of subcellular localization (WTAP commonly localizes to nuclear speckles; literature) when fixation/permeabilization conditions are optimized.
  • Chromatin/RNA-associated assays: Co-IP or RIP approaches to probe WTAP-containing ribonucleoprotein complexes (literature).

• Practical tips (general)

  • Confirm specificity with knockdown/knockout or peptide competition where feasible; this item is described as KD Validation grade in Product Data.
  • Include isotype controls and secondary‑only controls to evaluate background.
  • Optimize antibody dilution, incubation time, and antigen retrieval (for IHC) empirically; parameters are system‑dependent.

• Manufacturer Applications

  • Not specified in Product Data; no item‑specific tested applications are claimed here.

Reaction Conditions

• Applicability note

  • Classical reaction conditions (solvent, temperature, catalysts) for small‑molecule synthesis do not apply. Below are general, literature-based conditions for common immunoassay workflows; these are not item-specific tested parameters.

• General conditions (typical ranges; literature)

  • Western blot (WB): Primary incubation 1–16 h at 4–25°C in TBST + 1–5% BSA or milk; antibody dilution empirically determined (commonly 1:500–1:5,000 for many mAbs, but item-specific values are not provided).
  • Immunofluorescence (IF/ICC): Incubate 1–2 h at RT or overnight at 4°C in blocking buffer; permeabilization with 0.1–0.3% Triton X‑100 if nuclear targets are desired.
  • Immunoprecipitation (IP): 1–4 h capture at 4°C with gentle rotation; lysis buffers tailored to preserve complexes (e.g., 0.5% NP‑40, 150 mM NaCl) and protease/RNase inhibitors where relevant to WTAP complexes (literature).

• Controls and validation

  • Include knockdown/knockout controls, secondary‑only controls, and isotype controls to assess specificity and background.

• Caution

  • The above are general ranges; optimal conditions for this specific product are not specified and must be determined empirically.

Safety and Handling

• GHS and regulatory

  • GHS Classification / Signal Word / Pictograms / H‑Statements: Not specified for this item; refer to SDS.
  • As a protein research reagent, classification may vary by formulation (e.g., presence of sodium azide or other preservatives). Consult the product SDS for authoritative details.

• General laboratory precautions (antibody proteins; literature/general guidance)

  • PPE: Laboratory coat, gloves, and safety glasses. Avoid inhalation of aerosols and contact with skin/eyes.
  • Hygiene: Do not pipette by mouth. Wash hands after handling. Decontaminate work surfaces after use.
  • Incompatibilities: Avoid microbial contamination; keep containers tightly closed. Prevent exposure to strong oxidizers and proteases.
  • Sodium azide caution (if present; check SDS): Toxic; can form heavy‑metal azides in plumbing. Flush with copious water and collect waste appropriately.

• Handling and first aid (general)

  • Spills: Absorb with inert material; clean with appropriate detergent and water. For preserved formulations, treat as hazardous chemical waste per institutional policy.
  • Exposure: Rinse affected area with water. Seek medical attention if irritation persists. Provide SDS to healthcare personnel if needed.

• Special risks

  • Freeze–thaw damage: Repeated freeze–thaw cycles can cause aggregation and loss of activity—aliquot upon first thaw.
  • Biological origin materials: Although for research use only and typically non‑infectious, handle with standard biosafety level 1 practices unless otherwise specified.

Solvent Selection

• Applicability

  • As a proteinaceous reagent, this product is intended for use in aqueous buffers rather than organic solvents.

• Item-specific (from Product Data)

  • No formulation/buffer composition is provided; exact buffer, pH, and excipients are not specified for this item; refer to CoA/Spec Sheet.

• General/literature guidance for antibodies

  • Preferred buffers: PBS (10–50 mM phosphate, 100–150 mM NaCl, pH 7.2–7.6) or TBS for most immunoassays. Maintain near‑neutral pH.
  • Additives: 0.01–0.05% Tween‑20 can reduce nonspecific binding in wash buffers; 0.1–1% BSA or 1–5% nonfat milk for blocking (assay dependent). For storage, 10–50% glycerol may reduce freeze–thaw damage (verify assay compatibility).
  • Avoid: High percentages of organic solvents (e.g., acetonitrile, methanol, DMSO) which can denature antibodies; strong acids/bases; chaotropes unless intentionally used for SDS‑PAGE sample prep.

• Comparison (general)

  • PBS vs TBS: TBS may reduce background in phospho‑sensitive contexts due to phosphate interference; PBS is widely compatible and isotonic.
  • HEPES-buffered saline may offer improved pH stability near 7.4 for live‑cell compatible workflows.

Storage and Reconstitution

• Item-specific (from Product Data)

  • Storage Conditions: Store at −20°C. Avoid repeated freezing and thawing.
  • Shipping: Ice chest + ice pads.
  • Form/Appearance: Not specified for this item; refer to CoA/Spec Sheet.
  • Buffer composition / Preservatives / Concentration: Not specified for this item; refer to CoA/Spec Sheet.

• General handling guidance (literature; not item-specific)

  • Upon receipt, verify integrity, then centrifuge briefly to collect liquid from vial walls/cap before first use.
  • Aliquot immediately to single‑use portions to minimize freeze–thaw cycles; store tightly sealed to prevent evaporation and contamination.
  • If supplied liquid, maintain at −20°C (or as instructed); for short‑term (≤1–2 weeks), 2–8°C may be acceptable depending on formulation.
  • If supplied lyophilized (not specified here), reconstitute with sterile, neutral pH buffer (e.g., PBS) to the desired concentration using low‑bind tips/tubes. Allow to dissolve gently on ice; avoid vigorous vortexing.
  • For long‑term storage, many users add glycerol (final 10–50%) to liquid stocks; verify compatibility with intended assays.

• Stability note

  • Do not expose to elevated temperatures or prolonged light if conjugated. Track freeze/thaw history and avoid microbial contamination.

• Documentation

  • For definitive shelf life, reconstitution volume, and storage buffer, consult the CoA/Spec Sheet for this SKU.

Structure and Identity

Brief overview: This product is a mouse monoclonal antibody directed against the WTAP antigen. As a biomacromolecular reagent, it does not have a conventional small‑molecule structure.

• Item-specific (from Product Data)

  • SKU: Ab325644
  • Product Name: WTAP Mouse mAb
  • Type: Mouse monoclonal antibody (mAb) targeting WTAP (antigen designation inferred from product name)
  • CAS / CID / InChIKey / SMILES: Not specified for this item; refer to CoA/Spec Sheet.
  • Molecular Formula / Molecular Weight: Not specified for this item; refer to CoA/Spec Sheet.

• General/literature description of structure (for monoclonal IgG-class antibodies)

  • Typical mAbs are immunoglobulin G (IgG) proteins composed of two heavy and two light chains linked by disulfide bonds, forming a Y‑shaped heterotetramer with variable domains (VH/VL) that define antigen binding and constant domains that define isotype and effector functions.
  • Antibodies are glycoproteins; N‑linked glycosylation on the Fc region is common and can modulate stability and function.
  • The 2D “structure” can be described as domains: variable regions providing the paratope in complementarity‑determining regions (CDRs), and constant regions comprising CH1–CH3 (heavy) and CL (light) domains.
  • Typical overall size for IgG is ~150 kDa (literature), with a hydrodynamic diameter ~10–12 nm.

• Notes

  • Exact isotype, clonality details (clone ID), and any engineered modifications are not provided in the Product Data and should be confirmed on the CoA/Spec Sheet.

Synthetic Utility

• Not typically applicable

  • As a protein antibody reagent, this product is not used as a building block or reagent in classical small‑molecule synthesis. Instead, its value lies in analytical and affinity‑based applications (e.g., immunocapture, immunodetection).

• Related practical note (general)

  • For conjugation chemistry (e.g., labeling with fluorophores, HRP, or biotin), standard protein‑conjugation strategies can be applied (NHS‑ester to lysines, maleimide to cysteines) if permitted by the product’s formulation; however, the present item’s buffer composition and suitability for conjugation are not specified and should be confirmed before any modification.

Target Specificity

• Item-specific (from Product Data)

  • Antigen/Target: WTAP (Wilms Tumor 1‑Associating Protein) — inferred from the product name “WTAP Mouse mAb”.
  • Host/Type: Mouse monoclonal antibody (mAb) — from product name.
  • Clone ID: Not specified for this item; refer to CoA/Spec Sheet.
  • Isotype: Not specified for this item; refer to CoA/Spec Sheet.
  • Epitope mapping/Peptide immunogen: Not specified for this item; refer to CoA/Spec Sheet.
  • Species reactivity: Not specified for this item; refer to CoA/Spec Sheet.
  • Cross‑reactivity: Not specified for this item; refer to CoA/Spec Sheet.
  • Validation note: Grade indicates KD Validation (from Product Data), implying specificity supported by target knockdown studies.

• Usage note

  • For precise species reactivity, epitope, and cross‑reactivity data, consult the lot‑specific certificate or contact technical support.

Foire aux questions

What does KD Validation mean?

KD Validation indicates the product has been validated for use in rigorous antibody validation workflows. Validation may include positive/negative control samples, signal-to-noise testing, and protocol-specific QC. Lot-specific test data is published on the Certificate of Analysis where available.

How should this product be stored?

Store at ?20 °C. Avoid repeated freeze–thaw cycles. Aliquot into single-use volumes before freezing.

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.