Recombinant CDKN2A/p14ARF Antibody - Anticorps primaire, specific to CDKN2A, Rabbit IgG

Cat. No.: Ab095833

QUALITÉ & PURETÉ Recombinant, ExactAb™, Validated, See COA

Application

IF/ICC, WB

Host species

Lapin

Reactive species

Humain

Isotype

Rabbit IgG

Conjugation

Unconjugated

Vue d’ensemble

Rabbit anti Human CDKN2A/p14ARF Antibody, Recombinant, could be used for WB, ICC, IF and so on.
Application:
WB: 1/500-1/1000
ICC/IF: 1/50-1/200
Relevance
The gene for CDK2NA generates several transcripts/proteins which differ from each other in their first exons. Three of these transcripts are generated by alternative splicing (isoform 1 a.k.a p16INK4A, isoform 2 and isoform 3 a.k.a p12), two of which are known to function as inhibitors of CDK4 kinase. One other transcript that is generated from this gene contains an alternate reading frame (ARF), with the first exon located 20kb upstream of the remainder of the gene(isoform 4 a.k.a. p14ARF, p19ARF, ARF). In spite of the structural and some functional differences, all the proteins encoded by the CDKN2A gene are involved in cell cycle G1 control.

Spécifications

Product Name

Recombinant CDKN2A/p14ARF Antibody - Anticorps primaire, specific to CDKN2A, Rabbit IgG

Synonymes

ARF antibody|CDK4 inhibitor p16 INK4 antibody|CDK4I antibody|CDKN2 antibody|CDKN2A antibody|Cell cycle negative regulator beta antibody|CMM2 antibody|Cyclin dependent kinase 4 inhibitor A antibody|Cyclin dependent kinase inhibitor 2A antibody|INK4 antibody|INK4a antibody|Melanoma p16 inhibits CDK4 antibody|MLM antibody|MTS 1 antibody|MTS1 antibody|Multiple tumor suppressor 1 antibody|p14 antibody|p16 antibody|p16 INK4a antibody|p16INK4a antibody|p19 antibody|P19ARF antibody|TP16 antibody|CDKN2A|ARF|MLM|P14|P16|P19|CAI2|CMM2|INK4|MTS1|TP16|CDKN2|INK4A|MTS-1|P14ARF|P16INK4A

Spécifications et pureté

Recombinant, ExactAb™, Validated, See COA

Espèce hôte

Lapin

Spécificité

CDKN2A

Immunogène

A synthetic peptide derived from human CDKN2A/p14ARF (AA 75-132).

Contrôle positif

WB: HEK293 and LNCaP cell lysates. IF/ICC: HeLa cells.

Conjugation

Unconjugated

Grade

ExactAb™, Recombinant, Validated

Isotype

Rabbit IgG

Type de chaîne lumineuse

kappa

SDS-PAGE

150 kDa

Méthode de purification

Protein A purified

Concentration

See COA

Product Line

Produits biologiques

Stockage et expédition

Forme

Liquid

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 4°C short term (1-2 weeks). Store at -20°C long term (24 months). Upon receipt, it is recommended to aliquot. Avoid freeze/thaw cycle.

Images

Western Blot: Recombinant CDKN2A/p14ARF Antibody (Ab095833)

Recombinant CDKN2A/p14ARF Antibody (Ab095833) - Western Blot
All lanes: Recombinant CDKN2A/p14ARF Antibody (Ab095833) at 1/1000 dilution
Samples: Lysates at 20 µg per lane
Secondary: Goat Anti-Rabbit IgG H&L (HRP) at 1/20000 dilution
Predicted band size: 14 kDa
Observed band size: 16 kDa
Exposure time: 100.0 s

IF/ICC: Recombinant CDKN2A/p14ARF Antibody (Ab095833)

Recombinant CDKN2A/p14ARF Antibody (Ab095833) - IF/ICC
Immunofluorescent analysis of HeLa cells, using Recombinant CDKN2A/p14ARF Antibody (Ab095833) at 1/50 dilution.

Application

Application

Dilution info

IF/ICC

1/50-1/200

WB

1/500-1/1000

Cibles associées (humaines)

CDKN2A Tbio Tumor suppressor ARF (0 Activities)

Discover Target: CDKN2A

Activity Type	Activity Value -log(M)	Mechanism of Action	Activity Reference	Publications (PubMed IDs)

Application Protocols

• Item-specific tested applications, recommended dilutions, and positive controls: Not specified for this item; refer to CoA/Spec Sheet.

• General starting points (literature; adjust empirically and validate against controls):

  • Western blot (WB): 1:500 to 1:2,000 primary dilution; load 10–30 µg total protein per lane from cells with known ARF expression (e.g., oncogene-induced senescence models). Use anti-nucleolar markers for co-localization if needed.
  • IF/ICC: 1:200 to 1:800; fix with 4% PFA, permeabilize with 0.1–0.3% Triton X-100; include DAPI for nuclear counterstain.
  • IHC (FFPE): 1:100 to 1:500; perform HIER with citrate pH 6.0 or EDTA pH 9.0; include no-primary and isotype controls.
  • IP: 1–5 µg antibody per 500–1,000 µg lysate protein; rotate 1–4 h at 4°C; wash stringently.

• Controls and validation (literature):

  • Specificity: Use CDKN2A knockout/knockdown or locus-deleted lines as negative controls; confirm signal ablation.
  • Positive controls: Stimulate ARF expression via oncogenic stress (e.g., HRasV12) or use known ARF-positive cell lines/tissues.
  • Secondary antibodies: Match host/isotype precisely; verify cross-adsorption when working across species.

Biological Roles

Background on the target (literature; not product specifications):

• Gene/protein: CDKN2A produces multiple transcripts via alternative splicing and alternative reading frames. p14ARF (human; p19Arf in mouse) is a distinct protein from p16INK4a, sharing exons but encoded in an alternative reading frame.
• Primary function: p14ARF stabilizes p53 by antagonizing MDM2, sequestering MDM2 in the nucleolus and promoting p53-dependent transcriptional programs (cell cycle arrest, senescence, apoptosis depending on context). It acts as a tumor suppressor pathway component through the ARF–MDM2–p53 axis.
• Regulation: Induced by oncogenic stress (e.g., aberrant Myc, Ras); epigenetically silenced or deleted in many transformed contexts (CDKN2A locus deletions). Arf can also influence ribosomal biogenesis and nucleolar homeostasis.
• Localization: Predominantly nucleolar; can shuttle and interact with multiple nucleolar and nucleoplasmic proteins (e.g., NPM1).
• Molecular interactions: Binds MDM2; affects p53 stability; interacts with proteins involved in ribosome biogenesis and chromatin regulation (literature).

Relevance to this antibody:

• An anti-CDKN2A/p14ARF antibody enables detection, localization, and quantification of ARF protein in cell and tissue models to interrogate p53 pathway status, oncogene activation responses, and senescence-associated programs.

Item-specific note:

• Species reactivity, epitope, and validated applications for this SKU are not specified; consult the CoA/Spec Sheet for authoritative details.

Buffer Applications

While this product is an antibody rather than a buffering agent, correct buffer systems are critical for optimal assay performance.

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

• Practical buffer choices (general, literature-based):

  • Western blot: TBST (20 mM Tris, 150 mM NaCl, 0.1% Tween-20, pH 7.4–7.6). Blocking with 5% non-fat milk or 3–5% BSA. Primary incubation typically overnight at 4°C.
  • IHC (FFPE): Antigen retrieval in citrate buffer pH 6.0 or Tris-EDTA pH 9.0 (heat-induced). Washes in TBST or PBST. Use serum- or protein-based block compatible with secondary antibody host.
  • IF/ICC: PBS or HBSS-based buffers with 0.1–0.3% Triton X-100 for permeabilization (if intracellular antigen). Include 1–3% BSA for blocking.
  • IP: Lysis buffers matched to target localization; for nucleolar p14ARF, use gentle non-denaturing buffers (e.g., 20–50 mM Tris pH 7.5, 150 mM NaCl, 0.5% NP-40), plus protease inhibitors.

• Additive considerations:

  • Avoid sodium azide when using HRP-conjugated secondaries or live cells.
  • Include Mg2+/Ca2+ only if required; chelators (EDTA) can disrupt some protein interactions.

Green Alternatives

Environmental considerations for antibody reagents focus on formulation, packaging, and workflow rather than solvent replacement.

• Opportunities (general, literature-based):

  • Preservative strategy: Where feasible, use azide-free formulations to reduce acute aquatic toxicity; switch to sterile, single-use aliquots to control bioburden without chemical preservatives.
  • Packaging: Choose minimal, recyclable secondary packaging; request consolidated shipments to reduce cold-chain emissions.
  • Cold chain optimization: Glycerol-containing formulations can permit storage at −20°C (vs. −80°C), lowering energy use; validate stability before changing storage.
  • Wash buffers: Substitute nonionic detergents with lower environmental impact and use minimal effective concentrations; adopt closed-loop or on-demand buffer preparation to reduce waste.

• Trade-offs:

  • Preservative-free buffers demand stricter aseptic technique and single-use aliquots, increasing plastic consumption.
  • Higher glycerol content improves freeze–thaw resilience but raises COD in aqueous waste; balance against extended product lifetime.

• Item-specific note:

  • Exact formulation and preservatives for this SKU are not specified; refer to CoA/Spec Sheet before implementing green modifications.

Pharmaceutical Uses

This product is for research use only and is not intended for human or veterinary use, diagnostic procedures, or therapeutic applications.

• Item-specific regulatory status: Not specified for this item; refer to CoA/Spec Sheet. No GMP/Ph.Eur./USP grade claims are made in the provided Product Data.

• Relevant non-clinical applications (general):

  • In-process controls and QC in bioprocess development (e.g., monitoring p53 pathway markers in cell banks or engineered cell lines).
  • Exploratory biomarker research and preclinical mechanistic studies involving the ARF–MDM2–p53 axis.
  • Assay development: Antibody pairs (capture/detection) for non-clinical ELISAs after suitability and matrix interference testing.

• Formulation notes:

  • If the antibody contains preservatives (e.g., sodium azide), it is inherently incompatible with any in vivo use. For cell-based assays where preservatives are problematic, seek preservative-free or low-endotoxin formulations (not specified for this SKU; consult CoA/Spec Sheet).

Physical Properties

• Item-specific properties (from Product Data):

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

• General/literature characteristics for recombinant antibodies (for planning only; not product specifications):

  • If full-length IgG: typical MW ≈ 150 kDa (∼50 kDa heavy chain + ∼25 kDa light chain per monomer; non-reducing dimeric structure). Fragments: Fab ≈ 50 kDa; scFv ≈ 25–30 kDa.
  • Typical storage buffers: PBS or TBS, pH 7.2–7.5, often with 0.02–0.05% sodium azide and/or 10–50% glycerol for freeze–thaw protection (literature). Exact buffer for this item is not specified; refer to CoA/Spec Sheet.
  • Physical form: liquid solution or lyophilized cake; reconstitution details are item-specific (not specified here).
  • Stability: Antibodies are generally stable at 2–8°C short-term and at ≤−20°C long-term, with minimized freeze–thaw cycles; stability highly dependent on formulation (literature). Follow item-specific Storage Conditions below.

Quality and Grades

• Item-specific quality statements (from Product Data):

  • Grade/Purity: Recombinant, ExactAb™, Validated.
  • What this implies for this item:
    • Recombinant: Produced from a defined DNA sequence in a controlled expression system, enabling lot-to-lot sequence consistency and reduced batch variability compared with hybridoma-derived material.
    • ExactAb™: Brand indicator for sequence-defined, engineered antibodies with consistent performance characteristics; typically supports higher reproducibility in assays. Specific platform specifications for this SKU are not provided; refer to CoA/Spec Sheet.
    • Validated: The antibody has undergone in-house validation against CDKN2A/p14ARF. Exact tested applications, species reactivity, and validation data are not specified for this item; refer to CoA/Spec Sheet.

• General considerations for professional use (literature/guidance):

  • Validation hierarchy: Include specificity (knockdown/knockout controls), sensitivity (LOD/LOQ in assay), and reproducibility (lot bridging studies). Confirm epitope recognition status where known.
  • Purity expectations: Recombinant antibodies are usually affinity-purified (e.g., Protein A/G). Presence/absence of carrier proteins (BSA), stabilizers (glycerol, trehalose), and preservatives (azide) should be checked on the CoA for assay compatibility (e.g., mass spectrometry or live-cell imaging).

Reaction and Applications

Classical chemical reaction usage is not applicable to an antibody reagent. Instead, consider analytical/bioassay applications.

• Item-specific applications (from Product Data):

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

• General research applications for anti-CDKN2A/p14ARF antibodies (literature guidance):

  • Western blotting (WB): Detection of p14ARF (∼14–20 kDa apparent, isoform- and gel-condition-dependent) from cell lysates; nucleolar enrichment may be necessary depending on expression level.
  • Immunofluorescence (IF)/Immunocytochemistry (ICC): Visualization of nucleolar p14ARF; permeabilization conditions (e.g., 0.1–0.3% Triton X-100) and nucleolar counterstains can aid localization.
  • Immunohistochemistry (IHC-P): Analysis of FFPE sections for CDKN2A locus products; antigen retrieval conditions (e.g., citrate pH 6.0 or EDTA pH 9.0) must be optimized per tissue.
  • Immunoprecipitation (IP): Pull-down of p14ARF to study complexes with MDM2/p53 axis; stringent washes reduce nonspecific binding.
  • Knockout/siRNA controls: Validate specificity by loss-of-signal in CDKN2A-null backgrounds.

• Practical tips (literature):

  • Use fresh protease/phosphatase inhibitors during lysis.
  • Optimize primary antibody dilution by titration; begin with general ranges listed under Application Protocols and refine empirically.
  • Confirm species cross-reactivity with provided CoA or by sequence homology analysis of the immunogen region.

Reaction Conditions

Classical reaction conditions are not applicable. Below are general incubation parameters for common immunoassays employing anti-CDKN2A/p14ARF antibodies (literature guidance; NOT item specifications):

• Western blot (WB):

  • Sample: Denatured total lysate (RIPA or Laemmli), 10–30 µg per lane.
  • Primary incubation: Overnight at 4°C in TBST + 3% BSA.
  • Secondary: HRP-conjugated anti-IgG (host-specific), 1 h at RT.

• Immunofluorescence/ICC:

  • Fixation: 4% paraformaldehyde, 10–15 min; permeabilization: 0.1–0.3% Triton X-100, 5–10 min.
  • Primary: 1–2 h at RT or overnight at 4°C in PBS + 1–3% BSA.
  • Secondary: Fluorophore-conjugated anti-IgG, 1 h at RT, protect from light.

• IHC (FFPE):

  • Antigen retrieval: Citrate pH 6.0 or Tris-EDTA pH 9.0, 15–30 min (heat-induced); cool to RT.
  • Primary: 1–2 h at RT or overnight at 4°C.

• Immunoprecipitation (IP):

  • Incubation: 1–4 h at 4°C with rotation; capture with Protein A/G beads 1 h; wash with lysis buffer 3–5×.

Item-specific dilutions, host/isotype, and buffer composition are not specified for this item; refer to CoA/Spec Sheet and optimize empirically.

Safety and Handling

• Item-specific safety data (from Product Data):

  • GHS Classification, Signal Word, H-Statements, Pictograms: Not specified for this item; refer to SDS.
  • Research Use: For research use only.
  • Storage Conditions: Store at −20°C; avoid repeated freezing and thawing.
  • Shipping: Ice chest + ice pads (cold chain).

• General safety guidance for protein antibodies (literature/best practice; defer to SDS for authoritative instructions):

  • PPE: Use laboratory coat, appropriate gloves (nitrile), and eye protection when handling. Avoid ingestion, inhalation of aerosols, and contact with skin/eyes.
  • Biosafety: Treat all biological reagents as potentially hazardous. Handle under BSL-1 conditions unless local risk assessment indicates otherwise.
  • Incompatibilities: Avoid microbial contamination; do not mix with strong oxidizers or reactive chemicals that can denature proteins. Avoid metal-catalyzed oxidation; keep away from extreme pH (<5 or >9) and high temperature.
  • First aid (overview): If skin/eye contact occurs, rinse with water for 15 minutes; if inhaled as aerosol, move to fresh air; if ingested, rinse mouth and seek medical attention. Always consult the SDS.
  • Special risks: Repeated freeze–thaw can cause aggregation and loss of binding. Sodium azide (if present in buffer; not specified here) is toxic and may form explosive metal azides in plumbing—collect waste appropriately and avoid copper/lead drains.

Solvent Selection

This product is a protein antibody preparation and is not used as an organic solvent. Solvent selection in the chromatographic sense does not apply.

• Item-specific note:

  • Buffer composition for this SKU is not specified; refer to CoA/Spec Sheet.

• General laboratory guidance (literature) for antibody handling:

  • Preferred matrices: Aqueous buffers such as PBS or TBS at near-neutral pH (7.2–7.5).
  • Additives: For storage stability, cryoprotectants (e.g., 10–50% glycerol) and proteins (e.g., 0.1% BSA) may be included; verify compatibility with downstream assays. Preservatives like 0.02–0.05% sodium azide prevent microbial growth but are incompatible with HRP-based assays if not removed.
  • Avoid: Organic solvents (acetonitrile, methanol, DMSO) and strong detergents that can denature antibodies, unless specifically formulated as conjugates tolerant to such media.
  • For sample prep: Use compatible blocking buffers (e.g., 5% BSA or non-fat milk in TBST) during Western blot or IHC workflows to minimize nonspecific binding.

Storage and Reconstitution

• Item-specific instructions (from Product Data):

  • Storage Conditions: Store at −20°C; avoid repeated freezing and thawing.
  • Shipping: Ice chest + ice pads.
  • Reconstitution and formulation details: Not specified for this item; refer to CoA/Spec Sheet.

• General guidance (literature; follow item-specific CoA if provided):

  • If liquid: Aliquot upon receipt into single-use volumes to minimize freeze–thaw. Store at −20°C or −80°C depending on buffer composition (glycerol-containing formulations often stable at −20°C).
  • If lyophilized: Reconstitute with sterile water or buffer (e.g., PBS) to the desired concentration; gently invert—do not vortex vigorously. Allow 10–15 minutes for full dissolution on ice.
  • Additives: If compatible with intended use, include carrier protein (e.g., 0.1% BSA) and/or 10–50% glycerol for enhanced stability. Avoid sodium azide for live-cell work or peroxidase-based assays.
  • Storage stability: Short-term (≤1 week) at 2–8°C may be acceptable for working aliquots; for long-term, keep frozen. Protect from microbial contamination and light (for conjugated formats).
  • Documentation: Record lot number, aliquot concentrations, and freeze–thaw cycles. Always consult the CoA/SDS for authoritative stability and safety information.

Structure and Identity

Brief overview: This product is a recombinant antibody directed against human CDKN2A/p14ARF, supplied as an ExactAb™ reagent for research applications.

• Item-specific facts (from Product Data):

  • SKU: Ab095833
  • Product Name: Recombinant CDKN2A/p14ARF Antibody
  • Grade/Purity: Recombinant, ExactAb™, Validated
  • Category: Antibody (研究/实验用)
  • Molecular formula, InChIKey, SMILES, CAS, CID: Not applicable to protein biologics; not specified for this item; refer to CoA/Spec Sheet.

• Structural/biochemical identity (general, literature-based):

  • Target: CDKN2A locus product p14ARF (also known as ARF; human alternative reading frame product distinct from p16INK4a). Localizes predominantly to nucleolus; involved in p53 pathway regulation via MDM2 sequestration (literature).
  • Antibody architecture: Recombinant antibodies are commonly full-length IgG (≈150 kDa) or recombinant fragments (e.g., scFv, Fab; 25–55 kDa). The exact format, isotype, and species of this ExactAb™ are not specified for this item; refer to CoA/Spec Sheet.
  • Epitope/validation state: Validated (per Product Data) against CDKN2A/p14ARF; specific epitope mapping not specified for this item; refer to CoA/Spec Sheet.

• 2D/primary structure description (general):

  • Antibodies comprise two identical heavy chains and two identical light chains (if full-length IgG), with variable regions containing CDRs that confer antigen specificity; Fc region mediates effector functions. Recombinant production provides sequence-defined, batch-consistent material (literature).

Synthetic Utility

Not applicable. Antibodies are not used as reagents for chemical synthesis in the conventional sense.

• Related laboratory utility (general, literature-based):

  • Immunoaffinity purification: Antibody immobilized on a solid support can selectively capture p14ARF or its complexes from lysates for downstream proteomic analysis.
  • Analytical workflows: Use in WB, IHC, IF, IP, and proximity ligation assays for target-specific detection rather than chemical transformation.

• Item-specific note:

  • Details on coupling chemistries (e.g., amine coupling to resins) or antibody conjugation (biotin, HRP, fluorophores) are not specified for this item; refer to CoA/Spec Sheet or contact technical support for recommended protocols compatible with the ExactAb™ format.

Target Specificity

• Item-specific data (from Product Data):

  • Target/Antigen: CDKN2A/p14ARF.
  • Additional specificity details (clone, isotype, species reactivity, epitope, KD/affinity): Not specified for this item; refer to CoA/Spec Sheet.

• General background (literature):

  • CDKN2A/p14ARF is the human ARF isoform from the CDKN2A locus, functionally distinct from p16INK4a and primarily implicated in p53 pathway regulation via MDM2 interaction.
  • Because CDKN2A encodes multiple products, antibodies must be epitope-validated to distinguish p14ARF from p16INK4a; confirmation using isoform-specific genetic controls is recommended.

Foire aux questions

What does ExactAb? mean?

ExactAb? 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.

What does Recombinant mean?

Recombinant is a defined analytical purity tier suitable for workflows requiring a specific validated property. It specifies a minimum assay percentage, controlled limits on common impurities (water, heavy metals, residue on ignition), and lot-to-lot consistency. Recombinant is one of several purity tiers — choose based on the sensitivity and regulatory requirements of your method.

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.

Which species reactivity has this antibody been validated against?

Validated reactivity is listed on the PDP and on each lot's datasheet. We validate against the most commonly tested species for the target (Recombinant CDKN2A/p14ARF Antibody); cross-reactivity in untested species may exist but is not guaranteed. Contact technical support for specific species inquiries.