ARPC4 Antibody - Primary antibody, specific to ARPC4, Rabbit IgG

Cat. No.: Ab089682

GRADE & PURITY Carrier Free, ExactAb™, Azide Free, Validated, See COA

Application

IF/ICC, IHC, WB

Host species

Rabbit

Reactive species

Human, Mouse, Rat

Isotype

Rabbit IgG

Conjugation

Unconjugated

Vue d’ensemble

Rabbit anti Human ARPC4 Antibody, Polyclonal, could be used for WB, IHC and so on.
Application:

WB: 0.01-2µg/mL
IHC: 5-20µg/mL

IF/ICC: 2.3µg/mL
Protein Function
Functions as actin-binding component of the Arp2/3 complex which is involved in regulation of actin polymerization and together with an activating nucleation-promoting factor (NPF) mediates the formation of branched actin networks. Seems to contact the mother actin filament. 

Specifications

Product Name

ARPC4 Antibody - Primary antibody, specific to ARPC4, Rabbit IgG

Synonymes

Actin related protein 2/3 complex subunit 4 20 kD antibody | Actin related protein 2/3 complex subunit 4 20kDa antibody | Actin related protein 2/3 complex, 20-KD subunit antibody | Actin-related protein 2/3 complex subunit 4 antibody | ARC20 antibody | A

Spécifications et pureté

Carrier Free, ExactAb™, Azide Free, Validated, See COA

Espèce hôte

Rabbit

Spécificité

ARPC4

Immunogène

Recombinant ARPC4 expressed in E.coli (AA 3-165)

Contrôle positif

WB: HeLa, Jurkat, HEK293, THP-1, NIH/3T3 and C6 cell lysates. IHC: Human liver tissue. IF/ICC: HeLa cells.

Conjugation

Unconjugated

Grade

Azide Free, Carrier Free, ExactAb™, Validated

Isotype

Rabbit IgG

SDS-PAGE

150 kDa

Méthode de purification

Antigen affinity purified + 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℃ short term (1-2 weeks). Store at -20℃ long term (24 months). Upon receipt, it is recommended to aliquot. Avoid freeze/thaw cycle.

Images

Western Blot: ARPC4 Antibody (Ab089682)

ARPC4 Antibody (Ab089682) - Western Blot
All lanes: ARPC4 Antibody (Ab089682) at 1/1000 dilution
Samples: Lysates at 20 µg per lane
Secondary: Goat Anti-Rabbit IgG H&L (HRP) (Ab170144) at 1/20000 dilution

Predicted band size: 20 kDa
Observed band size: 19 kDa
Exposure time: 5.0 s

IHC: ARPC4 Antibody (Ab089682)

ARPC4 Antibody (Ab089682) - IHC
DAB staining on IHC
Samples: Human Liver Tissue
Primary Ab: 20 µg/mL ARPC4 Antibody (Ab089682)
Second Ab: 2 µg/mL HRP-Linked Caprine Anti-Rabbit IgG Polyclonal Antibody.

IHC: ARPC4 Antibody (Ab089682)

ARPC4 Antibody (Ab089682) - IHC
DAB staining on IHC
Sample: Human Liver Tissue
Primary Ab: 20 µg/mL ARPC4 Antibody (Ab089682)
Control: Used PBS instead of primary antibody
Second Ab: 2 µg/ml HRP-Linked Caprine Anti-Rabbit IgG Polyclonal Antibody. 

IF/ICC: ARPC4 Antibody (Ab089682)

ARPC4 Antibody (Ab089682) - IF/ICC
IF analysis of ARPC4 Antibody (Ab089682) in HeLa cells. The cells were fixed and permeabilized with 100% methanol for 5 minutes, and blocked with 5% BSA for 1 hour at room temperature. Cells were stained with ARPC4 Antibody (Ab089682) at 1/200 dilution in blocking buffer overnight at 4°C, and then incubated with Goat Anti-Rabbit IgG H&L (AF647) (Ab181958) at a dilution of 1/1000 for 1 hour at room temperature in the dark (red). Cells were counterstained with DAPI (blue).

Application

Application

Dilution info

IHC

5-20 µg/mL

WB

0.01-2 µg/mL

IF/ICC

2.3µg/mL

Application Protocols

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

General protocol outlines (for planning; not validated for this item)

• Western blot (WB):
  • Sample: Denatured lysates, 10–30 µg/lane. 12–15% SDS-PAGE for low-MW targets like ARPC4 (~20 kDa, literature).
  • Transfer: PVDF or nitrocellulose; confirm transfer of low-MW proteins by Ponceau.
  • Blocking: 5% BSA in TBS-T, 30–60 min.
  • Primary: Titrate antibody (e.g., 1:1,000–1:10,000 as a starting range, then optimize).
  • Secondary: HRP-conjugated anti-species IgG; ECL detection.
• Immunofluorescence (IF/ICC):
  • Fix: 4% PFA, 10 min; permeabilize 0.1% Triton X-100. Block with 1–3% BSA.
  • Primary: Incubate 1–2 h RT or overnight 4°C; include F-actin stain (e.g., phalloidin) for colocalization.
  • Secondary: Fluorophore-conjugated anti-species IgG; mount with antifade.
• Immunoprecipitation (IP/Co-IP):
  • Couple antibody to protein A/G resin; incubate clarified lysate 1–4 h at 4°C; wash with mild buffer; elute under gentle conditions for complex preservation.

Use these as starting points only. Determine optimal conditions empirically for your system and consult the item’s CoA for any product-specific instructions.

Biological Roles

About the antigen (general biology; not product specifications)

• ARPC4 (Actin Related Protein 2/3 Complex Subunit 4), also known as p20-ARC, is a core component of the heptameric Arp2/3 complex, which nucleates branched actin filament networks at the leading edge of motile cells (literature).
• Function: The Arp2/3 complex initiates new actin branches from the sides of pre-existing filaments, driving lamellipodial protrusion, endocytosis, vesicle trafficking, and pathogen motility. ARPC4 (together with ARPC2) forms the central scaffold that interacts with actin and nucleation-promoting factors (NPFs) such as WASP/Scar/WAVE family proteins.
• Regulation: Activation requires binding of NPF VCA domains, ATP, and actin monomers, with ARPC4 contributing to conformational transitions that position ARP2/ARP3 in a filament-like arrangement.
• Localization: Predominantly cytosolic with enrichment at the cell cortex and leading edge where actin branching activity is high (literature).
• Pathways: Involved in actin cytoskeleton organization, cell shape control, chemotaxis, and membrane remodeling. Genetic perturbation impacts cell migration and endocytic efficiency.
• Complex membership: Arp2/3 subunits include ACTR2 (ARP2), ACTR3 (ARP3), ARPC1A/B, ARPC2, ARPC3, ARPC4, and ARPC5/5L.

Utility of an ARPC4 antibody in biology (general)

• Enables detection/quantification of ARPC4 expression across cell types and perturbations (e.g., NPF activation, cytoskeletal drugs).
• Supports studies dissecting Arp2/3 assembly, complex stoichiometry, and dynamics via IP/Co-IP and microscopy colocalization with F-actin markers.

Note: The above describes antigen biology, not the performance characteristics of this specific antibody.

Buffer Applications

Applicability: Relevant as guidance for preparing and using antibody diluents and wash buffers. No item-specific buffer composition is provided.

Item-specific note

• Carrier free, azide free; exact buffer components and pH are not specified for this item; refer to CoA/Spec Sheet.

General buffer systems for antibody workflows (literature/practice)

• Storage buffer (short-term): PBS or TBS, pH 7.4, optionally with 10–50% glycerol for freeze–thaw protection if compatible with downstream use. For azide-free storage at 4°C, maintain sterility and consider 0.02–0.05% suitable preservative only if compatible with your application (not supplied in this product).
• Assay diluent: PBS/TBS with 0.05–0.1% Tween-20 and 0.1–1% BSA or casein to reduce nonspecific binding in WB, ELISA, and IF/ICC. For phospho-epitopes, BSA is preferred over milk.
• Wash buffer: PBS-T or TBS-T (0.05–0.1% Tween-20). Adjust ionic strength (150–500 mM NaCl) to tune stringency.
• Antigen retrieval (IHC-P): If using in paraffin sections, citrate buffer pH 6.0 or Tris-EDTA pH 9.0 are common retrieval buffers; optimize empirically (general guidance; not item-specific validation).
• Conjugation buffers: For NHS-ester coupling, use amine-free buffers (e.g., 50–100 mM sodium phosphate or carbonate, pH 8.3); for maleimide coupling, use phosphate or HEPES with 1–5 mM EDTA, pH 6.5–7.2.

Note: Validate buffer composition for your assay and consult the item’s CoA for any specific recommendations.

Green Alternatives

Context: Antibodies are aqueous protein reagents with minimal inherent environmental burden relative to many organic solvents, but cold-chain logistics and preservatives can impact sustainability.

Considerations and options (general; not item-specific specifications)

• Azide-free formulation: This product is azide free (item-specific), avoiding sodium azide toxicity and associated hazardous waste streams while remaining suitable for live-cell and enzyme-conjugation workflows.
• Shipping/storage optimization: Consolidate orders to reduce shipping frequency. If available, select lyophilized formats to enable refrigerated (rather than frozen) shipping; this item’s storage is -20°C (item-specific), so maintain cold-chain as provided.
• Buffer choices: Use phosphate- or Tris-based buffers prepared from concentrates to reduce packaging waste. Avoid unnecessary detergents and reduce Tween-20 concentration to the minimum effective level in washes.
• Reuse and waste reduction: Employ low-retention, reusable racks and validated cleaning protocols for gel boxes/blotting cassettes. Aliquot to minimize freeze–thaw–induced waste.
• Energy footprint: Plan experiments to minimize repeated freezer door openings; use inventory systems to shorten retrieval time. Where possible, store working aliquots at 4°C short-term to reduce freeze–thaw cycles (validate stability).

Trade-offs

• Preservatives vs safety: Omission of azide improves safety and conjugation compatibility but reduces antimicrobial protection; aseptic technique becomes more critical.
• Lyophilization vs liquid: Lyophilized antibodies can be more energy-efficient to ship/store but may require careful reconstitution and stabilization to maintain activity.

Pharmaceutical Uses

This product is an ARPC4 research antibody intended strictly for laboratory research use. It is not a drug substance, drug product, or excipient, and it is not intended for diagnostic or therapeutic applications.

• Pharmacopeial status: Not specified for this item; refer to CoA/Spec Sheet.
• Formulation role: Not applicable. Antibodies of this type are typically used as analytical reagents in discovery and quality-control research environments, not as formulation components.
• Manufacturing considerations: If used in bioprocess analytics (e.g., in-process testing), ensure that grade, documentation, and change control meet your internal QA requirements. This listing provides Carrier Free, Azide Free, Validated attributes; further QA/QC details are not specified here.

For any regulated use, obtain and review full documentation, including CoA, SDS, and any available validation reports. No clinical or diagnostic claims are made for this item.

Physical Properties

Item-specific properties

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Concentration: Not specified for this item; refer to CoA/Spec Sheet.
• Formulation: Carrier free; azide free. Specific buffer components, pH, and excipients are not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.

General/literature values for antibodies (for planning only; not product specifications)

• Typical IgG molecular mass: ~150 kDa; Fab ~50 kDa; scFv ~25–30 kDa (literature).
• UV absorbance: A280 ≈ 1.4 for 1 mg/mL IgG in PBS (literature approximation; sequence-dependent).
• Isoelectric point (pI): Often pI ~6–9 for mammalian IgG subclasses (broad literature range; sequence-dependent).
• Solubility: Antibodies are typically soluble in neutral aqueous buffers (e.g., PBS, TBS). Solubility and stability depend on sequence and formulation; avoid extreme pH and high ionic strength unless validated.
• Stability considerations: Repeated freeze–thaw can lead to aggregation and loss of binding activity; stabilizers (e.g., protein carriers, sugars) are sometimes used, but this item is carrier free (item-specific).

Note: Do not treat the above as specifications. Where exact numbers are required (e.g., concentration, extinction coefficient, buffer composition), consult the item’s CoA/Spec Sheet.

Quality and Grades

Item-specific quality attributes

• Carrier Free: The antibody solution contains no added protein carriers (e.g., BSA, gelatin). Advantages include lower background in immunoassays and compatibility with conjugation chemistries (e.g., HRP, biotin, fluorophores). Handle carefully to minimize nonspecific adsorption to plastics.
• Azide Free: No sodium azide preservative is present, facilitating use in live-cell applications and enzyme conjugations that are azide-sensitive. Note the reduced antimicrobial protection; maintain sterility and store as directed.
• ExactAb™: Typically denotes recombinant, sequence-defined production for lot-to-lot consistency and reproducible performance (brand-level description; consult CoA for specifics on clone/host/isotype).
• Validated: The item has been validation-tested by the manufacturer; specific validated applications, species reactivity, and performance metrics are not listed here. Refer to the CoA/Spec Sheet for assay-specific validation data.

What this means for your experiments

• Conjugation-ready: Carrier- and azide-free status is ideal for downstream labeling (NHS-ester dyes, maleimide linkers after partial reduction, click chemistry using azide-free protocols).
• Low background potential: Absence of stabilizer proteins reduces risk of carrier-derived interference in Western blot, IHC/IF, and IP.
• Documentation: For precise assay suitability (WB, IHC, IF, IP, FC), recommended dilutions, and antigen retrieval requirements, consult the item’s CoA/Spec Sheet.

Note: No item-specific purity %, endotoxin level, concentration, or isotype is provided here; Not specified for this item; refer to CoA/Spec Sheet.

Reaction and Applications

Item-specific applications

• Manufacturer Applications: Not specified for this item; refer to CoA/Spec Sheet for validated applications, species reactivity, recommended dilutions, and positive controls.

General use cases for an ARPC4 antibody (literature/practice; not a claim of performance for this specific lot)

• Western blot (WB): Detection of ARPC4 subunit of the Arp2/3 complex across cell/tissue lysates; expected apparent MW for ARPC4 is ~20 kDa (literature). Optimize blocking (e.g., 5% BSA) and adjust transfer conditions for low-MW proteins.
• Immunofluorescence (IF/IHC): Visualization of Arp2/3 complex localization in actin-rich structures (lamellipodia) with appropriate fixation (paraformaldehyde) and permeabilization (Triton X-100 or saponin). Epitope sensitivity to fixation may require optimization.
• Immunoprecipitation (IP/Co-IP): Enrichment of ARPC4 and associated Arp2/3 complex components (ARPC1–5, ARP2/ARP3) for downstream MS or WB.
• Flow cytometry (less common): Applicable if epitope is extracellularly accessible (ARPC4 is cytosolic; permeabilization required).
• Knockdown/overexpression validation: Antibody signal modulation following ARPC4 siRNA/CRISPR or cDNA overexpression supports specificity (control strategy).

Practical tips (general)

• Load controls: Use actin or GAPDH for WB; include peptide competition only if the antibody is peptide-derived and immunogen is available.
• Denaturation: For small proteins, use higher percentage gels (12–15%) or Tris–tricine systems to resolve bands sharpenly.
• Titration: Perform dilution series to identify optimal signal-to-noise; carrier-free format enables direct conjugation if desired.

Reaction Conditions

Not applicable in the small-molecule synthetic sense. However, for antibody-related chemistries and assays (general guidance; not item-specific specifications):

• Conjugation reactions: Perform NHS-ester labeling in amine-free buffers at pH ~8.3, 4–25°C for 30–120 min; quench with Tris or glycine. For maleimide-thiol coupling, partially reduce hinge disulfides with mild reducing agents (e.g., TCEP) and react at pH 6.5–7.2.
• Immunoassays: Typical incubation temperatures are 4°C (overnight) or room temperature (1–2 h) for binding steps; washing with PBS-T or TBS-T between steps. Optimize antibody dilutions empirically.
• Antigen retrieval (IHC-P): Heat-induced epitope retrieval in citrate pH 6.0 or Tris-EDTA pH 9.0, 15–30 min at near-boiling temperatures; validate conditions to preserve epitope integrity.

These are general literature/practice guidelines and are not validated conditions for this specific item. Consult the CoA/Spec Sheet for any product-specific recommendations.

Safety and Handling

GHS and hazard information (item-specific)

• Signal word: Not specified for this item; refer to SDS.
• H-statements / GHS classification / pictograms: Not specified for this item; refer to SDS.

General safety guidance for antibodies (informational; defer to SDS)

• Hazard profile: Protein aqueous solutions are generally considered low hazard. This product is azide free, eliminating sodium azide-related acute toxicity and metal/lead azide formation risks.
• Handling: Work using standard laboratory biosafety practices. Avoid creating aerosols. Use clean, low-binding plasticware when handling carrier-free proteins to minimize adsorption losses.
• PPE: Laboratory coat, gloves, and safety glasses are recommended. Use biological safety cabinet if handling in cell culture or if aerosols may be generated.
• Incompatibilities: Avoid strong oxidizers and proteases. Avoid contamination with microbes; use sterile technique for cell-based applications.
• First aid (general): If skin/eye contact occurs, rinse with water. If inhaled as aerosol, move to fresh air. If ingested, rinse mouth. Seek medical attention if irritation persists. Always follow institutional guidelines.
• Spill response: Absorb liquids with inert material; clean surfaces with suitable disinfectant (e.g., 70% ethanol) and water. Dispose according to local regulations and institutional biosafety procedures.
• Special risks: Repeated freeze–thaw can reduce performance due to aggregation; maintain cold chain and aliquot upon first thaw.

Always consult the official Safety Data Sheet (SDS) for authoritative guidance.

Solvent Selection

Applicability: As a protein reagent, “solvent” considerations pertain to aqueous buffer selection rather than organic solvent choice.

Item-specific facts

• Formulation: Carrier free, azide free; exact buffer composition not provided. Not specified for this item; refer to CoA/Spec Sheet.

General guidance for antibody diluents (literature/practice)

• Primary buffers: PBS (10–50 mM phosphate, 150 mM NaCl, pH 7.2–7.6) or TBS (Tris-buffered saline, pH 7.4–8.0) are standard for storage and assay dilutions.
• Additives to reduce nonspecific binding/adsorption: 0.05–0.1% Tween-20 for wash buffers; 0.1–1% BSA or appropriate blocking proteins for assay diluents. Since this product is carrier free, you may optionally add your preferred stabilizer immediately before use if compatible with your assay.
• Conjugation: For NHS-ester labeling, avoid primary amine-containing buffers (e.g., Tris, glycine). Use amine-free buffers (e.g., 50–100 mM sodium phosphate or carbonate, pH ~8.3). For maleimide-thiol coupling, ensure buffers are free of competing thiols (avoid DTT/βME) and use EDTA to chelate metals.
• Organic solvents: Avoid significant amounts of organic cosolvents; most antibodies lose activity in >10–20% organic. If required (e.g., for antigen retrieval or permeabilization), validate experimentally.

Comparison (general)

• PBS vs TBS: TBS can reduce background in phospho-protein blots where phosphate may interfere; PBS often preferred for physiological compatibility and cell-based work.

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.

General handling and reconstitution guidance (not item-specific specifications)

• Upon receipt: If provided frozen, keep at -20°C or colder until use. Thaw on ice. Mix gently by inversion—do not vortex vigorously.
• Aliquoting: Immediately prepare small, single-use aliquots in sterile, low-binding tubes to minimize freeze–thaw cycles and adsorption losses—especially important for carrier-free proteins.
• Reconstitution: If supplied lyophilized (not specified for this item), reconstitute with sterile, neutral pH buffer (e.g., PBS) to the desired concentration. Allow to dissolve gently on ice; avoid foaming.
• Optional stabilizers: If compatible with downstream applications, consider adding 10–50% glycerol for storage at -20°C and 0.1–1% carrier protein (e.g., BSA) to reduce surface adsorption. Do not add sodium azide if planning peroxidase-based assays or live-cell work.
• Short-term storage: Working aliquots may be kept at 4°C for several days if aseptic technique is maintained; monitor for microbial growth/turbidity.
• Freeze–thaw: Avoid repeated cycles. If necessary, snap-freeze aliquots (e.g., in a dry ice/ethanol bath) and store at -20°C or -80°C according to your lab’s stability findings.
• Documentation: For exact concentration, buffer composition, and any special stabilizers or lyophilization instructions, Not specified for this item; refer to CoA/Spec Sheet.

Always follow institutional biosafety procedures and consult the SDS for detailed safety information.

Structure and Identity

Overview: ARPC4 Antibody (SKU: Ab089682) is an affinity reagent directed against the ARPC4 antigen (Arp2/3 complex subunit 4). As an antibody product, it is a protein macromolecule rather than a small-molecule chemical. For research use only.

Item-specific facts (from Product Data)

• Product name: ARPC4 Antibody
• Grade/Purity: Carrier Free, ExactAb™, Azide Free, Validated
• Storage: Store at -20°C; avoid repeated freezing and thawing
• Shipped in: Ice chest + ice pads

Identity & structure (general antibody information; literature)

• Molecular class: Immunoglobulin (antibody) protein; exact isotype/host/clonality not specified for this item; refer to CoA/Spec Sheet.
• Molecular formula: Not applicable to proteins in a practical sense; item-specific formula not specified.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet. Typical IgG is ~150 kDa (literature), but recombinant fragments (Fab/scFv) can be smaller.
• Sequence/SMILES/InChIKey: Not applicable for antibodies; not provided for this item.
• Structural features: Antibodies commonly comprise variable regions (VH/VL) enabling antigen binding and constant regions (CH/CL) dictating effector/isotype properties. ExactAb™ typically denotes recombinantly defined sequence and lot-to-lot consistency (general).

2D/functional description (general)

• Two antigen-binding sites located in the variable domains recognizing epitopes within ARPC4. Without clone information, epitope location, paratope composition, and isotype are not specified for this item; refer to CoA/Spec Sheet.

Synthetic Utility

Not typically applicable. Antibodies are biological macromolecules used as affinity and detection reagents rather than as building blocks or reagents in chemical synthesis.

• If conjugation is planned (general guidance): The carrier-free, azide-free format is well-suited for chemical modification (e.g., NHS-ester dye labeling, SMCC-type heterobifunctional linkers, click-ready handles). Control degree of labeling (DOL) to preserve binding.
• For surface immobilization: Use protein A/G capture or directed coupling via Fc glycan oxidation and oxime ligation. Maintain native folding and avoid denaturing conditions.

No small-molecule synthetic transformations (e.g., coupling, reductions, protections) are relevant to this product’s typical use.

Target Specificity

Item-specific data

• Antigen/Target: ARPC4 (Arp2/3 complex subunit 4)
• Clone, isotype, host species, and epitope: Not specified for this item; refer to CoA/Spec Sheet.
• Species reactivity (tested): Not specified for this item; refer to CoA/Spec Sheet.
• Cross-reactivity: Not specified for this item; refer to CoA/Spec Sheet.

General notes (literature)

• ARPC4 is a conserved cytosolic protein; many anti-ARPC4 antibodies show cross-reactivity among human, mouse, and rat due to sequence conservation, but performance must be verified empirically for each species and application.

To confirm specificity for your system, use genetic controls (knockdown/knockout), peptide blocking (if immunogen is available), and orthogonal detection methods.