DTX41 Antibody

What is the relationship between DTX41, DTX1, and DTX4 proteins?

DTX1 (Deltex-1) is an approximately 75 kDa cytoplasmic and nuclear ubiquitin ligase that interacts with the Notch-1 intracellular domain and regulates Notch-induced gene transcription. It contains two WWE domains (amino acids 14-94 and 95-171) and one RING-type zinc finger (amino acids 411-472) . Within amino acids 1-147, human Deltex-1 shares 97% amino acid sequence identity with mouse and rat Deltex-1 . In plant research literature, QsDTX41 appears to be related to proanthocyanidin synthesis and programmed cell death pathways in cork oak .

What are the validated applications for DTX1/DTX4 antibodies in research?

Based on experimental validation, DTX1/DTX4 antibodies have demonstrated effectiveness in multiple applications:

These applications provide researchers with multiple methodological approaches to study DTX proteins in different experimental contexts .

How does DTX41 expression correlate with biological processes?

In cork oak research, QsDTX41 transcripts show significantly higher accumulation in both young-developed cork (YDC) (P = 0.0001) and trunk-derived cork (TDC) (P < 0.0001) compared to undifferentiated calli . Additionally, significantly higher expression was observed in TDC compared with YDC (P = 0.0002). This expression pattern correlates with other genes related to proanthocyanidin synthesis (QsLDOX, QsLAR, QsBAN) and appears to be associated with programmed cell death processes in cork formation .

What are the optimal conditions for preserving DTX1/DTX4 antibody activity?

For maximum antibody performance and longevity, researchers should follow these evidence-based storage protocols:

It is crucial to use a manual defrost freezer and avoid repeated freeze-thaw cycles that can compromise antibody performance .

What controls should be included when performing immunodetection of DTX1/DTX4?

To ensure experimental validity, incorporate these essential controls:

• Positive control lysates: Use validated cell lines such as K562 human chronic myelogenous leukemia cells or SW13 human adrenal cortex adenocarcinoma cells

• Negative controls: Include isotype control antibody (e.g., MAB0041) processed in parallel with DTX1/DTX4 antibody

• Secondary antibody controls: For Western blot, use HRP-conjugated Anti-Mouse IgG (HAF007); for immunofluorescence, use NorthernLights 557-conjugated Anti-Mouse IgG (NL007)

• Procedural controls: For flow cytometry, compare staining in permeabilized vs. non-permeabilized samples to verify intracellular localization

What protocol modifications are necessary for detecting DTX1/DTX4 in different cellular compartments?

Based on experimental evidence, researchers should consider these compartment-specific approaches:

• Cytoplasmic detection: Standard immunofluorescence with appropriate permeabilization has successfully detected DTX1/DTX4 in the cytoplasm of K562 cells

• Nuclear detection: Since DTX1 functions in both cytoplasmic and nuclear compartments, nuclear extraction protocols may be necessary to enrich for nuclear fractions

• Membrane association: For potential membrane-associated pools, subcellular fractionation followed by Western blotting can differentiate between soluble and membrane-bound populations

• Flow cytometry: For intracellular detection, fixation with paraformaldehyde followed by permeabilization with saponin has been validated for DTX1/DTX4 detection

How can DTX1/DTX4 antibodies be used to investigate Notch signaling pathways?

Researchers can employ several sophisticated approaches:

• Co-immunoprecipitation studies: Using DTX1/DTX4 antibodies to pull down protein complexes can identify interaction partners within the Notch pathway

• ChIP-seq analysis: Investigating potential roles of DTX1/DTX4 in transcriptional regulation by identifying genomic binding sites

• Proximity ligation assays: Visualizing direct protein-protein interactions between DTX1/DTX4 and Notch pathway components in situ

• Ubiquitination assays: Examining the E3 ligase activity of DTX1 on potential Notch pathway substrates

These approaches enable mechanistic investigation of DTX1/DTX4's role in regulating Notch-induced gene transcription .

What experimental techniques are effective for studying DTX41 in plant research models?

Based on cork oak research methodologies, several approaches have proven effective:

• Comparative gene expression analysis: qRT-PCR comparing QsDTX41 expression across different tissue types (calli, YDC, TDC) reveals developmental regulation

• Histochemical correlation: Vanillin-HCl staining for proanthocyanidins can be correlated with QsDTX41 expression to establish functional relationships

• Electron microscopy: Ultrastructural analysis of cellular changes during programmed cell death can be linked to DTX41 expression patterns

• Co-expression network analysis: Studying relationships between QsDTX41 and other genes involved in proanthocyanidin synthesis (QsLDOX, QsLAR, QsBAN) and programmed cell death (γVPE, ENDO4, PASPA3, RPN5A, XCP1)

What methodological approaches can distinguish between DTX41 and structurally similar proteins?

To ensure specificity in experimental detection:

• Epitope mapping: Characterize the specific epitope recognized by anti-DTX1/DTX4 antibodies to assess potential cross-reactivity

• Western blot validation: Confirm single-band detection at the expected molecular weight (~67 kDa for DTX1/DTX4) versus ~75 kDa for DDX41

• RNA interference: Use targeted siRNA knockdowns to validate antibody specificity through signal reduction

• Recombinant protein controls: Express tagged versions of DTX41 and similar proteins to establish detection specificities

• Orthogonal detection methods: Combine antibody-based detection with mass spectrometry for definitive protein identification

What are common challenges in DTX1/DTX4 detection and their solutions?

How should researchers interpret variable DTX1/DTX4 expression patterns in experimental models?

When analyzing DTX1/DTX4 expression data:

• Cell type-specific variation: Compare expression across multiple validated cell lines (K562, SW13) to establish baseline expectations

• Subcellular localization changes: Interpret cytoplasmic versus nuclear localization in the context of Notch pathway activation state

• Correlation with functional outcomes: Link expression patterns to downstream effects on Notch-induced gene transcription

• Developmental context: Consider temporal regulation, as seen in the progressive expression changes in cork development from calli to YDC to TDC

• Statistical validation: Apply appropriate statistical tests (as used in cork oak research with P-value thresholds) to determine significance of expression differences

What analytical approaches help distinguish specific from non-specific signals in DTX1/DTX4 detection?

To ensure signal specificity:

• Peptide competition assays: Pre-incubate antibody with immunizing peptide to confirm signal specificity

• Knockout/knockdown validation: Compare signal in wild-type versus DTX1/DTX4-depleted samples

• Multiple antibody validation: Use antibodies targeting different epitopes of DTX1/DTX4

• Signal quantification: Apply digital image analysis to quantify signal-to-noise ratios

• Physiological correlation: Verify that observed changes correlate with expected biological responses (e.g., Notch pathway modulation)

How can DTX1/DTX4 antibodies be used to investigate roles in cancer biology?

Building on detection in cancer cell lines (K562, SW13) , researchers can explore:

• Expression profiling: Compare DTX1/DTX4 levels across cancer types and correlate with clinical outcomes

• Functional studies: Investigate how DTX1/DTX4 modulation affects cancer cell proliferation, migration, and therapy response

• Pathway analysis: Examine crosstalk between Notch signaling and other oncogenic pathways mediated by DTX1/DTX4

• Biomarker potential: Evaluate DTX1/DTX4 as a potential diagnostic or prognostic biomarker in specific cancer types

• Therapeutic targeting: Explore ways to modulate DTX1/DTX4 activity for potential cancer treatment

What experimental approaches can reveal the molecular mechanisms of DTX41 in programmed cell death?

Based on cork oak research connecting QsDTX41 with programmed cell death :

• Time-course analysis: Monitor DTX41 expression changes throughout the progression of programmed cell death

• Genetic manipulation: Employ overexpression or silencing of DTX41 to assess effects on cell death pathways

• Protein interaction studies: Identify binding partners of DTX41 in cell death signaling networks

• Subcellular localization: Track changes in DTX41 distribution during cell death progression using fractionation and imaging

• Comparative systems biology: Compare DTX41 function in plant versus animal programmed cell death mechanisms

How might single-cell analysis technologies advance our understanding of DTX41/DTX1/DTX4 biology?

Emerging technologies offer new research possibilities:

• Single-cell RNA sequencing: Reveal cell-to-cell heterogeneity in DTX1/DTX4 expression within tissues

• CyTOF/mass cytometry: Simultaneously analyze DTX1/DTX4 expression alongside multiple signaling pathway components

• Spatial transcriptomics: Map DTX41 expression patterns within complex tissue architectures

• Live-cell imaging: Track dynamic changes in DTX1/DTX4 localization during cellular processes in real-time

• CRISPR screening: Identify genetic interactions with DTX1/DTX4 in specific cellular contexts