ADAP2 Antibody

Introduction to ADAP2 Antibody

ADAP2 antibodies are immunological reagents developed for the detection, quantification, and characterization of ArfGAP domain-containing protein 2 (ADAP2). These antibodies represent essential tools for researchers investigating the biological functions of ADAP2, particularly in the context of antiviral immunity and cellular trafficking pathways. Commercial ADAP2 antibodies are predominantly polyclonal in nature, derived from rabbit hosts, and are available in various formats including unconjugated and HRP-conjugated versions .

The development of highly specific ADAP2 antibodies has enabled significant advances in our understanding of the protein's role in cellular processes. These antibodies have been instrumental in revealing ADAP2's functions in membrane trafficking, macropinocytosis, and antiviral responses against RNA viruses such as dengue virus (DENV) and vesicular stomatitis virus (VSV) .

Applications of ADAP2 Antibodies in Research

ADAP2 antibodies have been employed in various experimental approaches to investigate the protein's biological functions. The primary applications include:

Western Blot Analysis

Western blotting represents a fundamental application of ADAP2 antibodies, enabling researchers to:

• Detect and quantify ADAP2 expression in various tissues and cell types

• Assess changes in ADAP2 expression levels following treatments such as interferon stimulation

• Examine post-translational modifications of ADAP2

The observed molecular weight of ADAP2 in Western blot experiments is approximately 44 kDa, consistent with its calculated molecular weight based on amino acid sequence .

Immunohistochemistry

Immunohistochemical applications of ADAP2 antibodies allow researchers to:

• Determine the tissue distribution of ADAP2 expression

• Examine the subcellular localization of ADAP2 in different tissue contexts

• Investigate changes in ADAP2 expression patterns in disease states

The Proteintech ADAP2 antibody (13706-1-AP) has been validated for immunohistochemistry, with positive detection in mouse spleen tissue . The recommended protocol for antigen retrieval involves TE buffer at pH 9.0, although citrate buffer at pH 6.0 may be used as an alternative .

ELISA Applications

ELISA-based detection using ADAP2 antibodies enables:

• Quantitative measurement of ADAP2 levels in biological samples

• High-throughput screening for changes in ADAP2 expression

• Evaluation of ADAP2 as a potential biomarker in various conditions

Multiple commercial ADAP2 antibodies have been validated for ELISA applications, including the HRP-conjugated antibody from Cusabio (CSB-PA889071LB01HU), which is specifically optimized for this technique .

ADAP2 Antibodies in Antiviral Research

ADAP2 antibodies have played a pivotal role in elucidating the protein's functions in antiviral immunity. Research utilizing these antibodies has revealed several important characteristics of ADAP2:

ADAP2 as an Interferon-Stimulated Gene

Studies employing ADAP2 antibodies have confirmed that ADAP2 is induced by type I interferons in a STAT1-dependent manner. Western blot analysis using these antibodies has demonstrated upregulation of ADAP2 expression following IFNβ treatment, similar to established interferon-stimulated genes such as the OAS family members .

Role in Restricting RNA Virus Replication

Immunofluorescence studies using ADAP2 antibodies have helped elucidate the protein's role in viral restriction. Key findings include:

• ADAP2 overexpression suppresses dengue virus (DENV) and vesicular stomatitis virus (VSV) infection in an Arf6 GAP activity-dependent manner

• ADAP2 has no consistent effect on coxsackievirus B (CVB) or Sendai virus (SeV) replication

• ADAP2 primarily restricts DENV infection at the stage of virion entry and/or intracellular trafficking

The following table summarizes experimental findings on ADAP2's antiviral effects:

Mechanism of ADAP2-Mediated Viral Restriction

Immunofluorescence microscopy using ADAP2 antibodies has revealed that:

• ADAP2 associates with actin-enriched membrane ruffles and induces pronounced membrane ruffling

• ADAP2 expression enhances macropinocytosis, as evidenced by increased dextran uptake

• ADAP2-containing cytoplasmic vesicles are enriched in dextran, a marker for fluid-phase uptake

• Incoming DENV and VSV particles associate with ADAP2 during their entry

• ADAP2 strongly associates with LAMP1-positive lysosomes but is excluded from EEA1- and Rab7-containing vesicles

These findings suggest that ADAP2 restricts viral infection by altering Arf6-mediated trafficking to disrupt viral entry and intracellular transport.

ADAP2's Role in Signal Transduction and Interferon Production

Beyond its direct antiviral functions, ADAP2 antibodies have facilitated the discovery of the protein's role in signal transduction pathways leading to interferon production. Research has identified ADAP2 as a key scaffolding protein in the RIG-I signaling pathway, which is critical for innate immune responses against RNA viruses .

Studies utilizing ADAP2 antibodies have revealed that:

• ADAP2 bridges the mitochondrial membrane-bound RIG-I adaptor MAVS with the downstream cytosolic complex of NEMO, TBK1, and IRF3

• This bridging function facilitates IRF3 phosphorylation, a critical step in type I interferon production

• ADAP2 independently orchestrates the interaction of TBK1 with NEMO and IRF3

• The ArfGAP domain of ADAP2 is essential and sufficient for mediating interferon responses

• TRAF3 acts as a trigger for ADAP2 to recruit RIG-I pathway component proteins into a single macromolecular complex

These findings highlight the multifaceted role of ADAP2 in antiviral immunity, encompassing both direct restriction of viral entry and facilitation of interferon-mediated antiviral signaling.

Technical Considerations for ADAP2 Antibody Applications

When utilizing ADAP2 antibodies for research applications, several technical considerations should be taken into account:

Dilution Recommendations

Different applications require specific antibody dilutions for optimal results:

Specificity Considerations

When selecting ADAP2 antibodies, researchers should consider:

• The specific epitope or region targeted by the antibody

• Cross-reactivity with orthologs from different species

• Potential cross-reactivity with related ArfGAP family proteins

• Validation data provided by the manufacturer