DDX19A Antibody

What is DDX19A and what are its key functions?

DDX19A (DEAD-box helicase 19A) belongs to the DEAD-box helicase family and functions as an ATP-dependent RNA helicase involved in mRNA export from the nucleus . It has been identified as a novel cytosolic RNA sensor that can activate the NLRP3 inflammasome during virus infection . Additionally, DDX19A is associated with NADPH oxidase 1 (NOX1)-mediated oxidative stress in tumor necrosis factor (TNF)-α-induced A549 cells . Its subcellular localization spans both the nucleus and cytoplasm .

What is the molecular structure of DDX19A?

DDX19A has two isoforms with molecular masses of 44 kDa and 54 kDa . The canonical protein has a reported length of 478 amino acid residues and a calculated molecular weight of 54 kDa . The gene is identified by NCBI Gene ID 55308 and UniProt ID Q9NUU7 .

What are common synonyms and alternative names for DDX19A?

DDX19A is also known as DDX19L, ATP-dependent RNA helicase DDX19A, DDX19-like protein, DEAD (Asp-Glu-Ala-As) box polypeptide 19A, DEAD (Asp-Glu-Ala-Asp) box polypeptide 19A, and DDX19-DDX19L .

What applications are DDX19A antibodies validated for?

DDX19A antibodies have been validated for multiple applications including Western Blot (WB), Immunohistochemistry (IHC), and ELISA . Some antibodies are available as part of matched antibody pairs for specialized applications like cytometric bead arrays .

What cell lines and tissues show positive expression of DDX19A?

Western blot analysis has confirmed DDX19A expression in HEK-293 cells, HepG2 cells, K-562 cells, and HeLa cells . For immunohistochemistry, human ovary tumor tissue has shown positive DDX19A detection . Notably, according to the Human Protein Atlas data, DDX19A protein expression is highest in cervical cancer among all common cancer types .

What antigen retrieval methods are recommended for IHC with DDX19A antibodies?

For immunohistochemistry applications using DDX19A antibodies, the suggested antigen retrieval method is TE buffer at pH 9.0 . Alternatively, antigen retrieval may be performed with citrate buffer at pH 6.0 . The choice of method may depend on the specific tissue type and fixation method.

How should DDX19A antibodies be stored and handled?

Storage recommendations vary by formulation. Standard antibody preparations should be stored at -20°C and remain stable for one year after shipment . For these preparations, aliquoting may be unnecessary for -20°C storage. For conjugation-ready antibodies in PBS-only formulations (without BSA and azide), storage at -80°C is recommended . Always refer to the specific product documentation for exact storage requirements.

How can researchers validate the specificity of DDX19A antibodies?

Validation strategies include:

• Testing in known positive cell lines (HEK-293, HepG2, K-562, HeLa)

• Performing knockdown/knockout experiments as negative controls

• Comparing observed molecular weights with expected values (44 kDa and 54 kDa for DDX19A)

• Comparing results across multiple applications (WB, IHC) when possible

• Verifying reactivity with human samples as documented in antibody specifications

How can DDX19A antibodies be used to study its role in cancer progression?

Research has shown that DDX19A is upregulated in cervical squamous cell carcinoma (CSCC) tissues and associated with metastasis and poor clinical outcomes . Researchers can employ DDX19A antibodies for:

• Immunohistochemical analysis of DDX19A expression in tumor tissues

• Correlation studies between DDX19A expression levels and clinical parameters

• Investigation of DDX19A's relationship with NOX1-mediated ROS production in cancer cells

• Monitoring DDX19A expression changes in metastatic versus non-metastatic samples

• Assessing DDX19A as a potential biomarker for cancer prognosis

What methods can be used to investigate the DDX19A-NOX1-ROS signaling axis?

To study this important pathway:

• Use Western blot with DDX19A antibodies to monitor expression changes in response to treatments

• Perform ROS detection assays in conjunction with DDX19A manipulation

• Design co-immunoprecipitation experiments to detect DDX19A-NOX1 interactions

• Conduct rescue experiments using NOX1 inhibitors or ROS scavengers in DDX19A-overexpressing cells

• Employ DDX19A antibodies in cell migration/invasion assays to correlate expression with functional outcomes

How can researchers study DDX19A's role in NLRP3 inflammasome activation?

Given DDX19A's function as an RNA sensor in inflammasome activation:

• Use DDX19A antibodies to track protein localization during viral infection

• Perform co-localization studies with NLRP3 components

• Design RNA-protein interaction studies to understand DDX19A's RNA sensing capabilities

• Monitor inflammasome activation markers in correlation with DDX19A expression levels

• Develop in vitro models combining DDX19A antibody-based detection with viral challenge experiments

Why might researchers observe multiple bands when using DDX19A antibodies in Western blot?

Multiple bands may appear due to:

• Detection of both known isoforms (44 kDa and 54 kDa)

• Post-translational modifications affecting protein migration

• Protein degradation during sample preparation

• Cross-reactivity with related DEAD-box helicases

• Alternative splicing producing additional variants

Researchers should compare observed bands with the expected molecular weights (44 kDa and 54 kDa) and consider using positive control samples with known DDX19A expression.

How can inconsistent DDX19A staining in IHC be addressed?

To improve IHC results:

• Optimize antigen retrieval conditions (try both recommended methods: TE buffer pH 9.0 and citrate buffer pH 6.0)

• Adjust antibody concentration within the recommended range (1:200-1:800)

• Extend incubation times to improve signal

• Use freshly prepared tissues with appropriate fixation

• Include positive controls (such as human ovary tumor tissue)

What controls are essential when studying DDX19A in metastasis models?

Critical controls include:

• DDX19A knockdown/overexpression validation by Western blot

• Comparison of multiple cell lines with different metastatic potentials

• In vivo metastasis models with appropriate controls (as described in research using SiHa cells)

• NOX1 inhibition controls to validate the DDX19A-NOX1-ROS pathway dependency

• Correlation with clinical parameters from patient samples to validate in vitro/in vivo findings

How might DDX19A serve as a therapeutic target or biomarker?

DDX19A shows promise as both a therapeutic target and biomarker based on recent findings:

• High expression of DDX19A correlates with lymph node metastasis and larger tumor size in CSCC

• Patients with high DDX19A expression have unfavorable prognoses compared to those with low expression

• Blocking the NOX1/ROS axis might serve as a potential therapeutic approach for patients with DDX19A-overexpressed CSCC

• DDX19A expression analysis could potentially help stratify patients for treatment decisions

What is known about DDX19A's role across different cancer types?

While DDX19A has been most extensively studied in cervical cancer, researchers should consider:

• Comparative expression analysis across cancer types (The Human Protein Atlas data suggests highest expression in cervical cancer)

• Investigation of DDX19A's role in other cancers with elevated ROS production

• Assessment of DDX19A as a pan-cancer metastasis regulator

• Correlation studies between DDX19A expression and treatment response in various cancer types

How can recombinant antibody technology improve DDX19A research?

Recombinant antibody technology offers several advantages:

• Unrivaled batch-to-batch consistency compared to traditional antibodies

• Easy scale-up for larger experiments

• Future security of supply through recombinant production

• Suitability for specialized applications including ELISAs, multiplex assays, mass cytometry, and multiplex imaging

• Compatibility with conjugation for custom applications