ADAM19 Antibody

What is ADAM19 and why is it important in research?

ADAM19, also known as a disintegrin and metallopeptidase domain 19 (meltrin beta), MLTNB, MADDAM, and FKSG34, is a protein encoded by the ADAM19 gene in humans . It has emerged as an important research target due to its association with various physiological processes and pathological conditions. The protein is structurally reported to be 105 kilodaltons in mass, and based on gene homology, orthologous proteins have been identified in canine, porcine, monkey, mouse, and rat models . ADAM19 has gained research significance particularly due to its consistent relation to pulmonary function in genome-wide association studies (GWAS) and its correlation with parameters of metabolic syndrome including BMI, relative fat, HOMA-IR, and triglycerides .

What experimental models are available for studying ADAM19 function?

Researchers have developed several experimental models to study ADAM19 function, with knockout mouse models being particularly valuable. The 2024 study created an Adam19 knockout (KO) mouse model by replacing exons 6 and 7 in Adam19 with a tdTomato red gene construct . This approach differs from previous models that targeted exons 10-12, which resulted in severe cardiac defects and early postnatal lethality . The newer model exhibits higher viability, making it more suitable for studying Adam19's role in adult physiology, particularly pulmonary function. Model validation can be performed using RNA sequencing (RNA-Seq) and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) . Additionally, diet-induced obesity mouse models have been used to investigate ADAM19's role in metabolic syndrome, allowing for interventional studies with neutralizing antibodies .

What are the commonly used applications for ADAM19 antibodies in research?

ADAM19 antibodies are utilized across multiple research applications. Based on the available commercial antibodies, the most commonly supported applications include:

• Western Blot (WB): For protein expression analysis

• Immunohistochemistry (IHC): For tissue localization studies

• Immunofluorescence (IF): For cellular and subcellular localization

• Flow Cytometry (FCM): For cell population analysis

• ELISA: For quantitative protein detection

• Immunoprecipitation (IP): For protein-protein interaction studies

How can gene targeting be optimized when creating ADAM19 knockout models?

Creating effective ADAM19 knockout models requires careful consideration of which exons to target. Earlier attempts targeting exons 10-12 resulted in severe cardiac defects with only 5% of mice surviving to postnatal day 8 . In contrast, the approach targeting exons 6 and 7 (replacing them with a tdTomato reporter construct) produced viable homozygous knockout mice suitable for adult phenotyping studies .

The methodology requires validation through:

• RNA-Seq analysis to confirm absence of functional transcripts

• RT-qPCR validation of knockout

• Sashimi plots for visualizing sequence coverage

• When possible, protein detection through western blot (though specific antibodies may be limiting)

When designing reporter constructs, researchers should consider potential protein misfolding issues - as noted in the 2024 study where tdTomato fluorescence could not be detected despite confirmation of tdTomato mRNA transcripts and protein expression .

What are the methodological considerations for using ADAM19 neutralizing antibodies in vivo?

When conducting in vivo studies with ADAM19 neutralizing antibodies, researchers should consider the following methodological aspects based on published protocols:

• Antibody administration timing: In the 2017 study, neutralizing antibodies were administered at week 10 of the dietary regimen, as ADAM19 protein increases at this time point in tissues such as liver .

• Dosage and frequency: The study protocol used 100 μg of antibody every 2 days via tail vein injection for a duration of two weeks .

• Control groups should include:

  • Standard diet control

  • High-fat diet with pre-immune IgG (as antibody control)

  • High-fat diet with ADAM19 neutralizing antibody

• Phenotypic assessment should include:

  • Body weight measurements (weekly)

  • Insulin tolerance tests (ITT; 0.5 U/kg)

  • Glucose tolerance tests (GTT; 1 g/kg)

  • Tissue collection for histological and molecular analysis

• Antibody characterization: Researchers should use well-characterized neutralizing antibodies with demonstrated efficacy, such as the pAb361 described in previous research .

What transcriptomic approaches provide insights into ADAM19 function?

Comprehensive transcriptomic analysis of ADAM19 function can be approached through:

• RNA-Seq analysis comparing wildtype and knockout tissues: This enables identification of differentially expressed genes and pathways affected by ADAM19 deletion .

• Bioinformatic processing pipeline:

  • Sequence quantification against reference transcriptomes (e.g., GENCODE vM33)

  • Quantification tools such as Salmon 1.10.0

  • Differential expression analysis using packages like limma-voom 3.54.2

  • Filtering criteria: false discovery rate (FDR)-adjusted p < 0.05, fold change ≥ 1.5, and group mean normalized transcript abundance ≥ 6 in at least one sample group

• Pathway analysis:

  • Gene set enrichment analysis (GSEA) to identify significantly enriched pathways (FDR p < 0.25)

  • Reference databases such as the Broad Molecular Signature Database (MSigDB)

• Human transcriptomic correlation studies: Large cohort studies like the San Antonio Family Heart Study (SAFHS) can be leveraged to correlate ADAM19 expression in peripheral blood mononuclear cells with metabolic parameters .

How does ADAM19 deficiency impact pulmonary function according to recent studies?

Recent research using Adam19 knockout mouse models has established a causal link between Adam19 deficiency and altered pulmonary function, supporting human GWAS findings . The 2024 study demonstrated that Adam19 knockout mice exhibit:

• Decreased pulmonary function parameters, validating human GWAS associations where ADAM19 gene variants are consistently related to pulmonary function.

• Altered immune cell profiles in response to challenges: When exposed to lipopolysaccharide (LPS), Adam19 knockout mice showed reduced responsiveness compared to wildtype mice, with 46% fewer neutrophils (p=0.032) and generally lower increases in total cell counts in bronchoalveolar lavage fluid (BALF) .

• These findings suggest that ADAM19 plays a role in regulating inflammatory responses in the lung, which may contribute to its effects on pulmonary function.

The methodology for assessing these impacts included pulmonary function testing in adult mice (aged 9-13 weeks) and immune cell differential analysis in BALF following LPS exposure .

What experimental approaches can assess ADAM19's role in lung inflammation?

To assess ADAM19's role in lung inflammation, researchers can employ several experimental approaches based on recent methodologies:

• LPS challenge model: Administer lipopolysaccharide to induce pulmonary inflammation in wildtype and Adam19 knockout mice .

• Cellular analysis of bronchoalveolar lavage fluid (BALF):

  • Total cell counting

  • Differential cell analysis focusing on neutrophils, macrophages, and lymphocytes

  • Statistical comparison between genotypes for inflammatory response magnitude

• Cytokine/chemokine profiling in lung tissue and BALF:

  • Though not explicitly mentioned in the provided studies for lung, the approach of measuring inflammatory mediators (such as TNF-α as done in liver tissue ) would provide valuable information about the inflammatory environment.

• Histopathological assessment:

  • Tissue sections stained for inflammatory markers

  • Quantification of inflammatory cell infiltration

• Functional assessment:

  • Pulmonary function testing to correlate inflammatory changes with functional outcomes

  • Measurement of airway hyperresponsiveness

These approaches would provide comprehensive assessment of how ADAM19 deficiency affects various aspects of pulmonary inflammation, from cellular recruitment to functional consequences.

How does ADAM19 expression correlate with metabolic syndrome parameters?

Research has established strong correlations between ADAM19 expression and various parameters of metabolic syndrome in both humans and animal models. In human studies using the San Antonio Family Heart Study cohort, ADAM19 expression in peripheral blood mononuclear cells strongly correlated with:

• BMI (Body Mass Index)

• Relative fat percentage

• HOMA-IR (Homeostatic Model Assessment for Insulin Resistance)

• Triglyceride levels

This indicates that ADAM19 expression may serve as a biomarker for metabolic syndrome. In animal models, ADAM19 expression was markedly increased in the liver and gonadal white adipose tissue of obese and type 2 diabetic mice . These findings suggest that ADAM19 expression not only correlates with but may functionally contribute to the development of obesity and insulin resistance.

What experimental designs best evaluate ADAM19 neutralization as a therapeutic approach for metabolic disorders?

Based on published research, an effective experimental design to evaluate ADAM19 neutralization for metabolic disorders would include:

• Animal model selection: Diet-induced obesity mouse model represents a physiologically relevant system that mirrors human metabolic syndrome development .

• Experimental groups (minimum requirements):

  • Control group: Standard diet control

  • Treatment control: High-fat diet with pre-immune IgG administration

  • Experimental group: High-fat diet with ADAM19 neutralizing antibody

• Treatment protocol:

  • Begin antibody administration when ADAM19 protein is elevated in metabolic tissues (week 10 in the referenced study)

  • Administer 100 μg antibody every 2 days via tail vein injection

  • Continue treatment for at least 2 weeks to observe metabolic effects

• Outcome measurements:

  • Body weight monitoring throughout the experiment

  • Insulin tolerance testing (ITT; 0.5 U/kg)

  • Glucose tolerance testing (GTT; 1 g/kg)

  • Inflammatory marker assessment (e.g., liver TNF-α levels)

  • Histopathological analysis of metabolic tissues

• Additional assessments that would strengthen the experimental design:

  • Blood lipid profile analysis

  • Energy expenditure measurements

  • Food intake monitoring

  • Analysis of adipose tissue inflammation

  • Assessment of hepatic steatosis

This comprehensive approach allows for thorough evaluation of ADAM19 neutralization on multiple aspects of metabolic syndrome.

What mechanisms link ADAM19 activity to insulin resistance development?

While the complete mechanistic pathway linking ADAM19 to insulin resistance is still being elucidated, current research suggests several potential mechanisms:

• Inflammatory modulation: ADAM19 neutralization reduced liver TNF-α levels in diet-induced obesity mouse models . Since TNF-α is a known contributor to insulin resistance, this suggests ADAM19 may promote insulin resistance through inflammatory pathways.

• Tissue-specific effects: ADAM19 expression is increased in both liver and gonadal white adipose tissue of obese and diabetic mice , suggesting it may have direct effects on insulin sensitivity in these metabolically active tissues.

• Clinical correlations: The strong correlation between ADAM19 expression and HOMA-IR in human studies further supports a functional link between ADAM19 activity and insulin resistance.

• Improvement with neutralization: The observation that neutralizing ADAM19 improves insulin sensitivity in diet-induced obesity models provides direct evidence of its role in insulin resistance development.

How can researchers address challenges with ADAM19 antibody specificity?

Researchers face challenges with ADAM19 antibody specificity, as noted in the 2024 study which mentioned a "lack of specific ADAM19 antibodies" . To address these challenges:

• Alternative validation approaches:

  • Gene expression analysis (RNA-Seq, RT-qPCR) to confirm target knockdown or overexpression

  • Reporter gene constructs (as used in the 2024 study )

  • CRISPR/Cas9-mediated tagging of endogenous ADAM19

• Antibody validation recommendations:

  • Test antibodies on knockout tissues as negative controls

  • Validate across multiple applications (WB, IHC, IF)

  • Compare results from antibodies targeting different epitopes

  • Pre-absorb antibodies with recombinant antigen to confirm specificity

• Application-specific considerations:

  • For Western blot: Include molecular weight controls and blocking peptides

  • For immunohistochemistry/immunofluorescence: Include isotype controls and knockout tissue sections

  • For functional studies: Compare multiple neutralizing antibodies targeting different domains

Since the commercially available ADAM19 antibodies come from 25 different suppliers with 156 total products , careful selection and validation are essential for experimental success.

What are the critical factors to consider when selecting ADAM19 antibodies for specific experimental applications?

When selecting ADAM19 antibodies for specific experimental applications, researchers should consider:

• Target epitope location:

  • Antibodies targeting different domains (metalloproteinase domain, disintegrin domain, cytoplasmic domain) may yield different results

  • Consider whether the application requires detection of full-length protein or processed forms

• Validation documentation:

  • Review manufacturer's validation data for your specific application

  • Check for citations in peer-reviewed publications using the antibody for your intended application

• Species cross-reactivity:

  • Ensure compatibility with your experimental model (human, mouse, rat, etc.)

  • Based on available products, there are antibodies reactive with human, mouse, and rat ADAM19

• Application-specific optimization:

  • For Western blot: Determine optimal concentration and blocking conditions

  • For IHC/IF: Optimize fixation and antigen retrieval methods

  • For neutralization studies: Test multiple concentrations to establish dose-response

• Conjugation requirements:

  • Choose between unconjugated antibodies or those with fluorescent tags (Alexa Fluor 488, Cy7) based on experimental design

  • Consider whether secondary detection or direct visualization is preferable

• Clone type considerations:

  • Monoclonal antibodies offer high specificity for a single epitope

  • Polyclonal antibodies may provide stronger signals through multiple epitope binding

What are the unexplored aspects of ADAM19 function that warrant further investigation?

Several unexplored aspects of ADAM19 function warrant further investigation:

• Substrate specificity:

  • Comprehensive identification of physiological substrates processed by ADAM19

  • Tissue-specific substrate profiles in pulmonary and metabolic tissues

• Regulatory mechanisms:

  • Factors controlling ADAM19 expression in different physiological and pathological states

  • Post-translational modifications affecting ADAM19 activity

• Tissue-specific functions:

  • Pulmonary-specific roles beyond those identified in the 2024 study

  • Cell type-specific functions within metabolically active tissues

• Signaling pathway integration:

  • Detailed mapping of how ADAM19 integrates with established inflammatory and metabolic signaling networks

  • Cross-talk between ADAM19-mediated pathways in different tissues

• Therapeutic potential expansion:

  • Investigation of ADAM19 in additional disease contexts beyond pulmonary function and metabolic syndrome

  • Development of small molecule inhibitors as alternatives to antibody-based targeting

• Structure-function relationships:

  • Crystal structure determination of ADAM19 domains

  • Structure-guided development of specific inhibitors

How might multi-omics approaches advance our understanding of ADAM19 biology?

Multi-omics approaches could significantly advance understanding of ADAM19 biology through:

• Integrated genomics, transcriptomics, and proteomics:

  • Correlate ADAM19 genetic variants with expression patterns and protein levels

  • Identify co-regulated gene networks across multiple tissues

• Degradomics:

  • Terminal amine isotopic labeling of substrates (TAILS) or other proteomics approaches to identify ADAM19 substrates

  • Comparison of the proteome and secretome between wildtype and Adam19 knockout tissues

• Metabolomics:

  • Profile metabolic changes in response to ADAM19 deficiency or neutralization

  • Correlate metabolite alterations with physiological phenotypes

• Single-cell multi-omics:

  • Characterize cell type-specific ADAM19 expression patterns

  • Identify cell populations most affected by ADAM19 deficiency

• Spatial transcriptomics/proteomics:

  • Map ADAM19 activity within tissue microenvironments

  • Correlate spatial expression patterns with local tissue functions

• Systems biology modeling:

  • Integrate multi-omics data to model ADAM19's role in complex physiological systems

  • Predict tissue-specific outcomes of ADAM19 modulation

These approaches would provide a comprehensive understanding of ADAM19 biology beyond what can be achieved through targeted studies.

What is the current consensus on ADAM19's role in health and disease?

Current research establishes ADAM19 as a multifunctional protein with significant roles in both development and disease. The consensus view based on available evidence indicates:

• Pulmonary function: ADAM19 is causally linked to pulmonary function, with deficiency affecting lung function and inflammatory responses .

• Metabolic regulation: ADAM19 exhibits pro-obesogenic and insulin resistance-enhancing properties, with its expression strongly correlating with metabolic syndrome parameters in humans and mice .

• Therapeutic potential: Neutralization of ADAM19 shows promise as a therapeutic approach for metabolic disorders, resulting in weight loss, improved insulin sensitivity, and reduced inflammation .

• Developmental roles: Earlier studies indicated ADAM19's importance in cardiac development, though the severity appears dependent on which exons are targeted in knockout models .

While these roles are becoming established, the molecular mechanisms underlying ADAM19's functions in different tissues remain incompletely understood.

What methodological advances are needed to overcome current limitations in ADAM19 research?

Several methodological advances would help overcome current limitations in ADAM19 research:

• Improved antibody development:

  • Generation of highly specific monoclonal antibodies for different ADAM19 domains

  • Development of application-specific validated antibodies

• Advanced genetic models:

  • Conditional and inducible knockout systems to study tissue-specific and temporal roles

  • Humanized mouse models expressing human ADAM19 variants

• Functional assay standardization:

  • Development of standardized activity assays for ADAM19 enzymatic function

  • Reporter systems for monitoring ADAM19 activity in real-time

• Therapeutic targeting:

  • Small molecule inhibitors with improved specificity over antibody approaches

  • Targeted delivery systems for tissue-specific ADAM19 modulation

• Translational methodologies:

  • Biomarker development for monitoring ADAM19 activity in human samples

  • Correlation of human genetic variants with functional outcomes