ARF6 Antibody - Anticorpo primario, specific to ARF6, Rabbit IgG

Cat. No.: Ab089477

QUALITÀ & PUREZZA Carrier Free, ExactAb™, Validated, 1.0 mg/mL

Application

IF/ICC, IHC, WB

Host species

Coniglio

Reactive species

Cat, consiglio, Maiale, Mouse, Umano, Zebrafish

Isotype

Rabbit IgG

Conjugation

Unconjugated

Panoramica

Rabbit anti Human ARF6 Antibody, Polyclonal, could be used for WB, IHC, ICC, IF and so on.
Application:
WB: 1/500-1/1000
IHC: 1/50-1/100
ICC/IF: 1/50-1/200
Protein Function：
GTP-binding protein involved in protein trafficking; regulates endocytic recycling and cytoskeleton remodeling. May modulate vesicle budding and uncoating within the Golgi apparatus. Functions as an allosteric activator of the cholera toxin catalytic subunit, an ADP-ribosyltransferase. Involved in the regulation of dendritic spine development (By similarity). Contributes to the regulation of dendritic branching and filopodia extension.

Specifiche

Product Name

ARF6 Antibody - Anticorpo primario, specific to ARF6, Rabbit IgG

Sinonimi

ADP ribosylation factor protein 6 antibody|ADP-ribosylation factor 6 antibody|ARF6 antibody|DKFZp564M0264 antibody|Small GTP binding protein antibody|Small GTPase antibody|ADP ribosylation factor 6 antibody

Specifiche e purezza

ExactAb™, Validated, Carrier Free, 1.0 mg/mL

Specie ospite

Coniglio

Specificità

ARF6

Immunogeno

A synthetic peptide derived from human ARF6

Controllo positivo

WB: Mouse liver, HeLa, MCF7, Hep G2, MDA-MB-231 and C6 lysates. IHC: Human lung cancer tissue. IF/ICC: C6 cells.

Coniugazione

Unconjugated

Grado

Carrier Free, ExactAb™, Validated

Isotipo

Rabbit IgG

SDS-PAGE

150 kDa

Metodo di purificazione

Immunogen affinity purified

Concentrazione

1.0 mg/mL

Product Line

Prodotti biologici

Stoccaggio e spedizione

Forma

Liquid

Condizioni di conservazione di stoccaggio

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

Spedito in

Ice chest + Ice pads

Stabilità e conservazione

Store at 4°C short term (1-2 weeks). Store at -20°C long term (12 months). Upon receipt, it is recommended to aliquot. Avoid freeze/thaw cycle.

Immagini

Western Blot: ARF6 Antibody (Ab089477)

ARF6 Antibody (Ab089477) - Western Blot
All lanes: ARF6 Antibody (Ab089477) at 1/1000 dilution
Samples: Lysates at 20 µg per lane
Secondary: Goat Anti-Rabbit IgG H&L (HRP) (Ab170144) at 1/40000 dilution

Predicted band size: 20 kDa
Observed band size: 18, 95 kDa
Exposure time: 25.0 s

IF/ICC: ARF6 Antibody (Ab089477)

ARF6 Antibody (Ab089477) - IF/ICC
Immunofluorescent analysis of ARF6 staining in C6 cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with ARF6 Antibody (Ab089477) in 3% BSA-PBS and incubated overnight at 4 °C in a hidified chamber. Cells were washed with PBST and incubated with a AF488-conjugated secondary antibody (green) in PBS at room temperature in the dark. Phalloidin -AF594 was used to stain Actin filaments (red). DAPI was used to stain the cell nuclei (blue).

IHC: ARF6 Antibody (Ab089477)

ARF6 Antibody (Ab089477) - IHC
Immunohistochemical analysis of ARF6 staining in human lung cancer formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with ARF6 Antibody (Ab089477) at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

Applicazione

Applicazione

Dilution info

IF/ICC

1/50-1/200

IHC

1/50 - 1/100

WB

1/500-1/1000

Obiettivi associati (umani)

ARF6 Tbio ADP-ribosylation factor 6 (0 Activities)

Discover Target: ARF6

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

Meccanismi d'azione

Application Protocols

Item-specific tested applications, recommended dilutions, incubation times, and positive control samples are Not specified for this item; refer to CoA/Spec Sheet.

General guidance (literature-based; adjust per CoA once available):

• WB: Start at 1:500–1:2,000 for primary incubation; adjust based on signal/background and membrane/blocking system. Use lysates known to express ARF6 (e.g., HeLa) as a positive control.
• IP: 1–5 µg antibody per 500–1,000 µg lysate protein; incubate 1–4 h at 4 °C; capture with Protein A/G beads.
• IF/ICC: 1:100–1:500 starting dilution; confirm fixation/permeabilization compatibility for ARF6.
• IHC: Optimize antigen retrieval (citrate pH 6 or Tris-EDTA pH 9); titrate antibody 1:100–1:1,000 depending on tissue and detection method.
• ELISA: If used as capture/detection after conjugation, begin with 0.5–2 µg/mL coating and 0.1–0.5 µg/mL detection.

Note: ExactAb™ and Carrier Free designations can facilitate custom conjugation; validate that conjugation does not impair antigen affinity.

Biological Roles

This antibody targets ARF6 (ADP-ribosylation factor 6), a small GTPase of the Ras superfamily.

• Core functions (general, literature):
  • Regulates endocytosis and fast recycling of membrane proteins to/from the plasma membrane, impacting integrin and receptor trafficking.
  • Orchestrates cortical actin cytoskeleton remodeling, influencing filopodia and membrane ruffling via effectors such as Rac1 signaling modules and PIP5-kinase-dependent phosphoinositide metabolism.
  • Coordinates membrane trafficking at the cleavage furrow during cytokinesis and participates in exosome/vesicle biogenesis.
• Molecular cycle (general):
  • Cycles between GDP-bound (inactive) and GTP-bound (active) states; activation is catalyzed by guanine nucleotide exchange factors (GEFs, e.g., ARNO/Cytohesin family) and inactivation by GTPase-activating proteins (GAPs).
  • Post-translational modifications (e.g., myristoylation at the N-terminus, general for ARF family) enable membrane association.
• Localization (general): Predominantly plasma membrane and endosomal compartments; redistributes dynamically in response to signaling cues.
• Research relevance: Antibodies against ARF6 enable detection and quantification of its expression/localization, mapping of trafficking pathways, and assessment of regulatory perturbations by GEFs/GAPs or small-molecule modulators.

Note: The above biological information is general literature knowledge about ARF6. Item-specific species reactivity, epitope, and performance characteristics for this antibody are not specified and should be confirmed on the CoA/Spec Sheet.

Buffer Applications

• Applicability: Highly relevant. Antibody performance depends on buffer composition, pH, ionic strength, and additives. Item-specific buffer supplied with this product is not specified; refer to CoA/Spec Sheet.

• Common working buffers (general, literature):

  • PBS (10 mM phosphate, 137 mM NaCl, 2.7 mM KCl, pH 7.2–7.6) or TBS (20 mM Tris, 150 mM NaCl, pH 7.4–7.6) for dilution and storage.
  • Add 0.01–0.05% Tween-20 for wash steps in WB/ELISA/IHC to reduce nonspecific binding.
  • Include 0.1–5% BSA or nonfat dry milk for blocking in WB/ELISA; avoid carriers in conjugation workflows (product is Carrier Free—add blockers only in assay steps, not to the stock vial, unless compatible).
  • For IF/ICC, use permeabilization buffers (e.g., PBS + 0.1–0.3% Triton X-100 or 0.05–0.1% saponin) as appropriate for ARF6 intracellular detection.

• pH ranges (general): Most IgG antibodies are stable near neutral pH (6.8–7.6). Avoid low ionic strength for long storage to minimize aggregation.

• Storage additives (post-aliquoting, general): 30–50% glycerol at −20 °C can reduce freeze–thaw damage; 0.02–0.05% sodium azide may be used as a preservative for non-cell-based applications (verify compatibility and safety). Do not add azide if using HRP conjugates or living cells.

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

Green Alternatives

• Solvent and reagent stewardship (general for immunoassays):
  • Prefer aqueous buffers (PBS/TBS) and minimize use of organic solvents. When blocking/washing, use biodegradable surfactants where feasible and at the lowest effective concentrations.
  • Replace methanol-based antigen retrieval or fixation where possible with buffered formaldehyde or glyoxal-based systems; assess epitope compatibility.
  • Use reusable staining trays and reduce single-use plastics; pre-block plastics to minimize adsorption losses rather than increasing protein concentration.
  • Substitute chromogenic substrates that generate hazardous waste with fluorescent detection when compatible with instruments.
• Cold chain efficiency: Consolidate shipments and use high-efficiency coolants to reduce packaging waste; store promptly to avoid product loss.
• Antibody format considerations:
  • Carrier-free preparations reduce extraneous protein load in waste streams and improve conjugation efficiency, minimizing additional purification steps.
  • Where possible, use recombinant, sequence-defined antibodies (often denoted by proprietary marks such as ExactAb™) to enhance reproducibility and reduce animal-derived variability.
• Trade-offs: Some greener fixes (e.g., methanol-free permeabilization) may alter epitope exposure; always re-optimize conditions to balance sustainability with data quality.

Pharmaceutical Uses

This is a research-use-only antibody reagent and is not intended for human or veterinary use in therapeutics, diagnostics, or as a drug substance/excipient.

• Pharmacopeia/compendial status: Not applicable; no pharmacopeial monograph is indicated.
• Formulation roles: Not applicable—this antibody is supplied for laboratory research workflows (e.g., analytical immunoassays) rather than for pharmaceutical formulation.
• Manufacturing notes: If used in bioprocess analytics (in-process testing or product characterization), ensure the reagent is qualified internally. Carrier-free format is advantageous when users require custom conjugation for assay development.

Any clinical or therapeutic applications are outside the scope of this product. For GMP-grade or diagnostic-grade reagents, consult appropriate suppliers and regulatory guidance.

Physical Properties

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Grade/Purity: Carrier Free; ExactAb™; Validated (see Quality & Grades tab)
• Solubility: Antibodies are water-soluble in physiological buffers (general, literature). Item-specific buffer composition is not specified; refer to CoA/Spec Sheet.
• pI/pKa, logP, refractive index: Not applicable/not specified for this item.
• Typical protein attributes (general, literature):
  • Antibodies are amphipathic glycoproteins; stability depends on pH (commonly stable near neutral), ionic strength, and temperature.
  • Aggregation can occur upon freeze–thaw or at low ionic strength; gentle handling and appropriate buffers mitigate this.
• Boiling/melting points, vapor pressure, density: Not applicable to macromolecular aqueous protein formulations.
• Concentration: Not specified for this item; refer to CoA/Spec Sheet.
• Additives/stabilizers: Carrier Free designation implies no added protein carriers such as BSA (see Quality & Grades); any other stabilizers are not specified for this item and should be confirmed on the CoA/Spec Sheet.

Quality and Grades

• Item-specific designations (from Product Data):
  • Carrier Free: Supplied without protein carriers such as BSA, gelatin, or casein. This is advantageous for conjugation (e.g., biotin/fluorophore/enzymes), surface immobilization, or sensitive assays where carriers interfere. It also enables accurate protein quantitation without carrier background. Handle gently to avoid adsorption losses; consider adding your own stabilizer post-aliquoting if compatible with downstream use.
  • ExactAb™: A proprietary designation. In common usage, such marks denote sequence-defined, recombinant, or otherwise highly standardized antibodies that emphasize lot-to-lot consistency and reproducibility. Specific attributes for this item (e.g., recombinant origin, sequence) are not specified; consult the CoA/Spec Sheet for definitive details.
  • Validated: Indicates this antibody has undergone internal performance checks. Exact validation matrices/applications, species reactivity, and methods are not specified for this item; refer to CoA/Spec Sheet.
• Purity/grade caveats:
  • No titer, purity %, endotoxin, or host/isotype information is provided here; these are Not specified for this item; refer to CoA/Spec Sheet.
• Practical implications:
  • For conjugation workflows, the carrier-free format reduces pre-cleanup and improves labeling efficiency.
  • For quantitative immunoassays, absence of carriers can reduce background but may increase adsorption to plastics; pre-block tubes/tips or add a compatible inert protein after aliquoting if adsorption is observed.

Reaction and Applications

As an analytical antibody reagent, the concept of chemical “reactions” is not directly applicable. Instead, this product is used in immunochemical applications that rely on selective antigen recognition.

• Typical research applications (general, literature):
  • Western blotting (WB) to detect ARF6 in denatured lysates; requires compatibility with reducing/denaturing conditions.
  • Immunoprecipitation (IP)/co-IP to enrich ARF6 and associated complexes from cell extracts.
  • Immunofluorescence (IF)/immunocytochemistry (ICC) for subcellular localization of ARF6.
  • Immunohistochemistry (IHC) on fixed tissues where ARF6 expression is profiled; epitope retrieval may be required depending on fixation.
  • Flow cytometry (FC) if ARF6 is probed intracellularly following permeabilization.
  • ELISA or sandwich immunoassays when paired with suitable capture/detection reagents.
• Target biology context (general, literature): ARF6 is a small GTPase regulating endocytic recycling, actin remodeling, and plasma membrane dynamics. Antibodies against ARF6 are often employed to study GTP/GDP cycling, localization, and pathway modulation by GEFs/GAPs.
• Practical tips (general):
  • Validate species reactivity and recommended dilutions with the item’s CoA before use.
  • Optimize fixation (paraformaldehyde vs methanol) and permeabilization conditions (saponin vs Triton X-100) for intracellular targets like ARF6.
  • Include appropriate positive controls (cells known to express ARF6) and negative controls (isotype control or knockout/siRNA samples).

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

Reaction Conditions

Classical reaction conditions do not apply. For immunochemical workflows, consider the following general, literature-based conditions and optimization parameters:

• Western blotting: 1–5% blocking agent (milk or BSA) in TBS-T/PBS-T; antibody incubation typically 1–16 h at 4–25 °C; washing with TBS-T/PBS-T. Reducing/denaturing sample prep (SDS, DTT/βME) as required. Optimize antibody dilution empirically.
• Immunoprecipitation: Lysis buffers (e.g., NP-40, Triton X-100, or RIPA) at 4 °C with protease inhibitors; incubate 1–4 h with antibody, then capture with Protein A/G beads; wash stringently; elute for SDS-PAGE or MS.
• IF/ICC: Fix with 3–4% paraformaldehyde (or methanol if compatible); permeabilize; block with serum or BSA; incubate with primary and appropriate labeled secondary; image under fluorescence microscopy.
• IHC: Formalin-fixed paraffin-embedded tissue may require citrate or EDTA antigen retrieval; optimize retrieval pH/temperature empirically.
• ELISA: Coating 0.5–2 µg/mL capture antibody; blocking with protein-based buffer; incubations at room temperature or 37 °C as assay dictates.

Item-specific incubation times, temperatures, and dilutions are Not specified for this item; refer to CoA/Spec Sheet and optimize per sample type.

Safety and Handling

• GHS classification, signal word, pictograms, H-statements: Not specified for this item; refer to the SDS for authoritative safety information.
• General laboratory handling (proteins/antibodies):
  • Wear appropriate PPE (lab coat, gloves, safety glasses). Avoid generating aerosols; handle solutions in a biological safety cabinet if working with biological samples.
  • Avoid repeated freeze–thaw cycles to minimize aggregation and loss of activity.
  • Use only for research purposes. Not for human or animal therapeutic or diagnostic use.
• Storage incompatibilities: Avoid strong oxidizers and extremes of pH; do not mix with proteases. Keep away from microbial contamination; use sterile technique.
• First-aid overview (general): If skin or eye contact occurs, rinse with water. If inhaled as aerosol, move to fresh air. If ingested, rinse mouth with water. Seek medical attention as needed and present SDS.
• Spills and decontamination: Wipe with absorbent material and clean area with a suitable disinfectant (e.g., 70% ethanol); dispose of waste in accordance with institutional biosafety procedures.
• Waste: Protein solutions are typically nonhazardous chemical waste but may be biohazardous if mixed with biological samples; follow local regulations.
• Shipping: Shipped in ice chest with ice pads as provided in Product Data to maintain cold chain.

Solvent Selection

This product is an aqueous, protein-based antibody reagent. Traditional organic solvent selection guidance does not apply.

• Applicability: Not typically applicable to antibodies. Use physiological, protein-compatible buffers.
• Recommended media (general, literature):
  • Neutral pH buffers such as PBS or TBS for routine handling.
  • Add 0.01–0.05% nonionic detergent (e.g., Tween-20) to reduce nonspecific binding in washes (assay dependent).
  • For long-term storage after aliquoting, many labs add 30–50% glycerol (v/v) at −20 °C to reduce freezing damage (ensure compatibility with intended applications).
• Avoid: High percentages of organic solvents, extremes of pH, strong denaturants, or chaotropes unless deliberately performing denaturation assays.
• See also: Buffer Applications and Storage & Reconstitution tabs for practical guidance.

Storage and Reconstitution

• Storage conditions (from Product Data): Store at −20 °C. Avoid repeated freezing and thawing.
• Shipping: Supplied in an ice chest with ice pads to maintain the cold chain.
• Reconstitution: Not specified for this item; refer to CoA/Spec Sheet. If lyophilized, reconstitute gently in sterile, neutral pH buffer (e.g., PBS) to the desired concentration; avoid vigorous agitation.
• Aliquoting: Upon first thaw/reconstitution, aliquot into low-protein-binding tubes to avoid repeated freeze–thaw cycles. Keep on ice during use.
• Stabilizers (user-added, optional and assay-dependent): For storage after aliquoting, many labs add 30–50% glycerol (v/v) and store at −20 °C to prevent freezing. Preservatives such as 0.02–0.05% sodium azide can be added for stock solutions not intended for live-cell applications (verify compatibility and safety). Do not add carriers if you plan downstream conjugation; this product is Carrier Free.
• Short-term handling: Store at 2–8 °C for up to a few days during intensive use; return to −20 °C for longer-term storage. Protect from microbial contamination.
• Freeze–thaw guidance: Minimize cycles; thaw on ice, mix gently by inversion, and briefly spin down to collect contents.

Item-specific concentration, buffer composition, and format (liquid vs lyophilized): Not specified for this item; refer to CoA/Spec Sheet.

Structure and Identity

• Item type: Research-grade antibody reagent targeting ADP-ribosylation factor 6 (ARF6)
• SKU: Ab089477; Product Name: ARF6 Antibody
• CAS: Not applicable to antibodies
• Molecular formula: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet. (General note: IgG-class antibodies are typically ~150 kDa in literature; actual for this item not specified.)
• SMILES/InChIKey: Not applicable to proteins/antibodies
• Structural features (general, literature):
  • Antibodies are Y-shaped immunoglobulin proteins composed of two heavy and two light chains, stabilized by disulfide bonds, with variable domains forming the antigen-binding sites (paratopes).
  • If monoclonal and recombinant, the binding site is sequence-defined; for polyclonal mixtures, multiple epitopes may be recognized. Status for this item is not specified; refer to CoA/Spec Sheet.
• Target description (general, literature): ARF6 is a small GTPase (Ras superfamily) associated with the plasma membrane and endosomal system, cycling between GDP-bound (inactive) and GTP-bound (active) states.
• Epitope/clone/host/isotype: Not specified for this item; refer to CoA/Spec Sheet.
• 2D description in words: Not a small molecule; an immunoglobulin macromolecule with variable (Fab) domains that bind ARF6 and a constant (Fc) domain mediating effector interactions. Exact sequence/format for this item is not specified.

Synthetic Utility

Not applicable as a chemical building block or reagent. This product is a protein antibody used for biochemical detection and affinity-based assays rather than for driving or participating in chemical synthesis.

• Relevant laboratory utilities (general):
  • Affinity enrichment of ARF6 from complex mixtures (immunoprecipitation) prior to downstream proteomics or immunoblotting.
  • As a capture or detection reagent in immunoassays designed to quantify ARF6.
  • Potential use in proximity assays (e.g., PLA) or imaging workflows after user-conjugation to enzymes or fluorophores.

No named organic reactions, catalysts, or retrosynthetic applications apply.

Target Specificity

• Antigen/Target: ARF6 (ADP-ribosylation factor 6)
• Clone, epitope, host species, isotype, and species reactivity: Not specified for this item; refer to CoA/Spec Sheet.
• Cross-reactivity and blocking peptide availability: Not specified for this item; refer to CoA/Spec Sheet.
• Validation status: Labeled as “Validated” in Product Data; specific validation details (applications/samples) are not provided here.

For definitive specificity data, consult the item’s CoA/Spec Sheet and SDS.