DTX21 Antibody

What is DTX2 protein and what are its key characteristics?

DTX2 (deltex homolog 2) is a protein originally identified in Drosophila with human homologs. The human DTX2 gene (Gene ID: 113878) encodes a protein with calculated molecular weights of either 253aa/28 kDa or 575aa/64 kDa, though Western blot detection typically observes it at approximately 68 kDa . This discrepancy suggests potential post-translational modifications or alternative splicing events that researchers should consider when interpreting their results. DTX2's UNIPROT ID is Q86UW9, and the protein has been studied in multiple human cell lines including HeLa, HepG2, MCF-7, and A549 cells .

What types of DTX2 antibodies are currently available for research?

Current research tools include both polyclonal and monoclonal antibodies targeting DTX2. The polyclonal antibody (e.g., 17615-1-AP) is derived from rabbit IgG, while the monoclonal antibody (e.g., 67209-1-Ig) comes from mouse IgG2a . These antibodies differ in their applications and species reactivity: the polyclonal antibody demonstrates reactivity with human, mouse, and rat samples, making it suitable for cross-species studies, while the monoclonal antibody has validated reactivity specifically with human samples, offering potentially higher specificity for human-focused research .

How do researchers select between polyclonal and monoclonal DTX2 antibodies?

Selection should be based on experimental requirements and the biological question being investigated. Polyclonal DTX2 antibodies offer broader epitope recognition and cross-species reactivity (human, mouse, rat), making them valuable for detection of DTX2 across different experimental models . Monoclonal antibodies provide higher specificity to a single epitope, potentially reducing background and cross-reactivity, but with more limited species range (primarily human samples) . For novel applications or detection in non-validated species, researchers should perform validation studies before proceeding with extensive experiments.

What are the proper storage and handling protocols for DTX2 antibodies?

DTX2 antibodies should be stored at -20°C in their recommended buffer solution (PBS with 0.02% sodium azide and 50% glycerol, pH 7.3) . These storage conditions ensure stability for approximately one year after shipment. For the polyclonal antibody (17615-1-AP), aliquoting is noted as unnecessary for -20°C storage . For the monoclonal antibody (67209-1-Ig), similar conditions apply, though researchers may want to aliquot larger volumes to minimize freeze-thaw cycles that could degrade antibody quality over time . Some preparations may contain 0.1% BSA for additional stability .

How should researchers validate the specificity of DTX2 antibodies in their experimental systems?

Validation should include positive controls using cell lines known to express DTX2 (HeLa, HepG2, MCF-7, or A549 cells) . Include negative controls such as samples where DTX2 is absent or knocked down. Compare the observed molecular weight (approximately 68 kDa) with expected size ranges. When investigating novel systems or species not explicitly validated in product documentation, preliminary experiments using multiple detection methods (such as combining the polyclonal and monoclonal antibodies to target different epitopes) can provide stronger evidence of specificity and reduce the risk of misinterpreted results.

What approaches can resolve discrepancies between calculated and observed molecular weights of DTX2?

The discrepancy between calculated (28 kDa/64 kDa) and observed (68 kDa) molecular weights of DTX2 is significant and requires careful experimental design to address . To investigate this phenomenon, researchers should consider implementing:

• Treatment with various deglycosylation enzymes to identify potential glycosylation contributions

• Phosphatase treatments to detect phosphorylation events affecting migration

• Analysis of various tissues/cell types to identify tissue-specific post-translational modifications

• Alternative detection methods such as mass spectrometry to confirm protein identity

• Cloning and expression of different isoforms to compare migration patterns

How can researchers optimize immunoprecipitation protocols for DTX2 studies?

While specific immunoprecipitation (IP) protocols for DTX2 require optimization for each research context, general recommendations include:

• For monoclonal antibody-based IP, using protein G-conjugated beads typically yields better results than protein A

• Starting with 1-5 μg of antibody per 500 μg of total protein lysate and adjusting based on results

• Including protease and phosphatase inhibitors in lysis buffers to preserve protein integrity

• Performing pre-clearing steps with beads alone to reduce non-specific binding

• Validating IP results with Western blot using a separate DTX2 antibody recognizing a different epitope

• For co-immunoprecipitation studies, optimizing crosslinking conditions depending on the strength of the interaction being studied

What are key considerations when designing immunofluorescence experiments with DTX2 antibodies?

While specific validation for immunofluorescence applications is not directly addressed in the provided data, researchers planning immunofluorescence studies should:

• Begin with fixation optimization, testing both paraformaldehyde and methanol-based protocols

• Include antigen retrieval steps if working with tissue sections

• Titrate antibody concentrations, starting with ranges recommended for Western blot, then adjusting

• Include co-localization studies with known subcellular markers to confirm expected distribution patterns

• Validate specificity using siRNA knockdown or genetic knockout controls

• Consider species cross-reactivity when designing experiments in non-human models, as the monoclonal antibody has only been validated in human samples

What controls should be incorporated in experimental designs involving DTX2 antibodies?

A robust experimental design should include multiple controls:

• Positive controls: Lysates from validated cell lines (HeLa, HepG2, MCF-7, or A549)

• Negative controls: DTX2 knockout/knockdown samples or tissues/cells known not to express DTX2

• Loading controls: When performing Western blot, include standard loading controls such as GAPDH or β-actin; the GAPDH (14C10) Rabbit mAb has been cited as one of the top antibodies in research publications

• Secondary antibody controls: Samples incubated with secondary antibody only to identify non-specific binding

• Peptide competition: If available, competing the antibody with immunizing peptide can confirm specificity

How should researchers approach DTX2 studies in disease models?

When investigating DTX2 in disease contexts, researchers should consider:

• Baseline characterization: Establish normal expression patterns in relevant tissues using both antibodies (polyclonal for broader detection, monoclonal for higher specificity)

• Paired sample design: Compare DTX2 expression in matched normal/disease samples from the same individuals when possible

• Multiple detection methods: Combine protein detection (Western blot) with mRNA analysis (qPCR) to distinguish between transcriptional and post-transcriptional regulatory effects

• Isoform-specific analysis: Given the multiple calculated molecular weights (28 kDa/64 kDa) , design experiments to distinguish between potential isoforms or processed forms

• Functional validation: Complement expression studies with functional assays after modulating DTX2 levels

What emerging technologies should researchers consider for advanced DTX2 studies?

Building on recent advancements in antibody technology, researchers investigating DTX2 might benefit from:

• AI-designed antibodies: New tools like RFdiffusion can help design antibodies with improved specificity for challenging epitopes

• Single-cell analysis: Combining DTX2 antibodies with single-cell technologies to understand heterogeneity within populations

• CRISPR-based validation: Using CRISPR/Cas9 to create knockout controls that definitively validate antibody specificity

• Nanobody and scFv approaches: Smaller antibody fragments may access epitopes that conventional antibodies cannot reach

• Multiplexed imaging: Combining DTX2 detection with other markers to understand pathway interactions and cellular context

How does DTX2 relate to other members of the deltex protein family?

While the search results don't provide specific information on DTX2's relationship to other deltex family members, researchers should consider:

• Comparative expression analysis of multiple deltex family members across tissues

• Using both DTX2-specific antibodies in combination with antibodies against related family members to investigate potential functional redundancy or compensation

• Cross-reactivity testing to ensure antibodies specifically detect DTX2 and not related proteins with similar epitopes

• Evolutionary analysis of conserved domains that may inform functional studies

What considerations should guide the development of next-generation DTX2 research tools?

Based on current trends in antibody development and research needs, future DTX2-focused tools might benefit from:

• Recombinant antibody technology: Recombinant monoclonal antibodies represent about a quarter of the most popular antibody products and offer improved consistency

• Isoform-specific antibodies: Developing antibodies that can distinguish between the different potential DTX2 isoforms (28 kDa vs. 64 kDa)

• Conjugated antibodies: Direct fluorophore or enzyme conjugation to reduce protocol steps and background

• Humanized versions: For therapeutic applications or in vivo studies in humanized models

• Antibody engineering using computational approaches like those described for RFdiffusion could lead to highly optimized DTX2-binding proteins