DTX47 Antibody

What is DTX47 and what biological systems express this protein?

DTX47 is a protein found in Arabidopsis thaliana (Mouse-ear cress), a model organism widely used in plant biology research. The commercially available DTX47 antibody specifically targets this plant protein with confirmed reactivity to Arabidopsis thaliana samples . This protein belongs to the broader DTX family, which in other organisms like humans includes members involved in important signaling pathways.

What are the physical properties of the DTX47 Antibody?

The DTX47 Antibody is supplied as a liquid preparation containing 50% glycerol with 0.01M PBS (pH 7.4) and 0.03% Proclin 300 as a preservative . It is a polyclonal antibody raised in rabbits against recombinant Arabidopsis thaliana DTX47 protein . The polyclonal nature provides recognition of multiple epitopes, potentially increasing detection sensitivity compared to monoclonal alternatives.

What applications has the DTX47 Antibody been validated for?

The antibody has been specifically tested and validated for ELISA and Western Blot applications for the identification of the antigen . These applications represent fundamental techniques in protein expression analysis that can be incorporated into broader experimental workflows for studying plant protein function.

How should experimental controls be designed when using DTX47 Antibody?

When designing experiments with DTX47 Antibody, researchers should incorporate appropriate positive and negative controls. Similar to experimental designs used with other plant antibodies, positive controls should include samples known to express DTX47, while negative controls might include samples from DTX47 knockout lines or non-Arabidopsis plant species. This approach mirrors experimental design principles demonstrated in studies of other specialized antibodies .

What are the recommended storage and handling protocols for DTX47 Antibody?

The DTX47 Antibody should be stored at -20°C or -80°C upon receipt . Researchers should avoid repeated freeze-thaw cycles to maintain antibody integrity and functional activity . This storage recommendation aligns with general best practices for antibody preservation, where temperature stability is critical for maintaining epitope recognition capabilities.

How can specificity of the DTX47 Antibody be validated in experimental systems?

Antibody specificity validation should follow rigorous protocols including:

What sample preparation methods optimize DTX47 detection in plant tissues?

Optimal sample preparation for plant tissue analysis with DTX47 Antibody likely requires:

• Effective tissue disruption using appropriate buffers

• Inclusion of protease inhibitors to prevent degradation

• Optimization of protein extraction conditions specific to membrane-associated proteins

• Careful quantification of total protein before immunoassay application
  While specific optimization for DTX47 would require empirical testing, these approaches represent standard methodological considerations for plant protein analysis.

How does epitope accessibility influence DTX47 detection in different experimental conditions?

The accessibility of epitopes can significantly impact antibody-antigen interactions. For DTX47 Antibody, which was developed against the recombinant protein , native protein conformations in different extraction conditions may affect epitope accessibility. This consideration parallels observations with other antibodies, such as those against diphtheria toxin mutants, where single amino acid substitutions can dramatically alter antibody recognition patterns .

What are the optimal blocking conditions for Western blot applications with DTX47 Antibody?

Optimization of blocking conditions typically requires empirical testing. For plant proteins like DTX47, researchers should consider:

• Testing both BSA and non-fat dry milk as blocking agents at different concentrations (3-5%)

• Evaluating Tween-20 concentrations (0.05-0.1%) in wash buffers

• Determining optimal antibody dilution ranges to maximize signal-to-noise ratio

• Testing different incubation temperatures and durations
  These variables should be systematically evaluated to develop an optimized protocol specific to DTX47 detection.

What are common causes of inconsistent results when using DTX47 Antibody in Western blot?

Inconsistent Western blot results may stem from several factors:

How can researchers differentiate between specific binding and non-specific interactions?

Distinguishing specific binding from background requires multiple controls:

• Include a no-primary antibody control to assess secondary antibody specificity

• Perform peptide competition assays where excess immunizing peptide should abolish specific signal

• Compare patterns between wild-type and knockout/knockdown samples

• Evaluate signal persistence across multiple independent experiments
  These approaches allow researchers to confidently differentiate specific DTX47 detection from experimental artifacts.

What analytical methods are recommended for quantifying DTX47 expression levels?

For quantitative analysis of DTX47 expression, researchers should:

• Use appropriate normalization controls (housekeeping proteins)

• Apply densitometry software with standardized analysis parameters

• Establish a standard curve with recombinant protein when absolute quantification is required

• Consider multiple biological and technical replicates for statistical validity
  These analytical approaches align with established quantitative western blot methodologies used in implementation science research settings .

How does DTX47 compare functionally to other DTX family proteins?

While DTX47 is specific to plant systems, other DTX family proteins like DTX4 in humans function as E3 ubiquitin ligases involved in Notch signaling and neurogenesis . DTX4 has also been implicated in the innate immune response, where it participates in regulating type I interferon signaling . Comparative studies between plant DTX47 and mammalian DTX proteins could reveal evolutionary conservation of functional domains and mechanisms.

What experimental approaches would best investigate DTX47's role in plant stress responses?

Investigation of DTX47 in plant stress responses might employ:

• Expression analysis under various abiotic and biotic stress conditions

• Generation of transgenic lines with altered DTX47 expression

• Co-immunoprecipitation studies to identify interaction partners

• Subcellular localization studies under different environmental conditions
  These experimental approaches would provide complementary data on DTX47's functional role in plant biology.

What emerging technologies might enhance DTX47 research in the future?

Emerging technologies with potential application to DTX47 research include:

• CRISPR-Cas9 gene editing for precise modification of DTX47 in Arabidopsis

• Advanced imaging techniques for in vivo monitoring of DTX47 localization

• Proteomics approaches to identify the complete DTX47 interactome

• Single-cell analysis to reveal cell-type specific expression patterns
  These methodological advances represent the cutting edge of implementation science in biological research .