ATX4 Antibody
Description
Arabidopsis ATX4 in Plant Biology
ATX4 (Arabidopsis thaliana trithorax-like 4) is a plant-specific histone methyltransferase belonging to the trithorax-group (TrxG) proteins. It regulates gene expression by catalyzing histone H3 lysine 4 methylation (H3K4me), a modification associated with transcriptional activation.
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Function: ATX4 and its homolog ATX5 negatively regulate drought stress responses in Arabidopsis by modulating abscisic acid (ABA) signaling pathways .
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Mutant Studies:
Relevant Data:
No studies in the provided sources describe the development or use of antibodies targeting Arabidopsis ATX4. Research on these proteins relies on genetic mutants and transcript analysis rather than antibody-based tools.
Potential Misinterpretation of "ATX4"
The term "ATX" is also associated with other proteins, but these are unrelated to Arabidopsis ATX4:
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Autotaxin (ATX/ENPP2): A lysophospholipase D enzyme involved in immune cell migration . Antibodies against ATX have been studied for their role in blocking lymphocyte homing , but this protein is distinct from Arabidopsis ATX4.
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ACKR4: Atypical chemokine receptor 4 in mice, for which monoclonal antibodies (e.g., A4Mab-1) have been developed . These antibodies target ACKR4, not ATX4.
Antibody Development for Related Targets
While ATX4-specific antibodies are undocumented, the search results highlight methodologies for antibody generation against other targets:
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Anti-tau Antibodies:
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Anti-ACKR4 Antibodies:
Technical Insights into Antibody Characterization
The provided sources emphasize critical parameters for antibody validation:
Product Specs
Composition: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Target Background
Q&A
Frequently Asked Questions on ATX4 Antibody in Academic Research
What experimental controls are critical when studying ATX4’s role in ABA signaling?
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Negative controls: Include atx4/atx5 double mutants to account for functional redundancy .
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ABA treatment controls: Use WT seedlings treated with ABA to benchmark H3K4me3 dynamics at stress-response genes (e.g., RD29A, ABI5) .
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Histone modification controls: Pair ATX4 ChIP with H3K4me3/H3K4me2 quantification to distinguish catalytic activity from non-enzymatic roles .
How can I resolve contradictions in ATX4 antibody performance across plant tissues?
What advanced techniques enhance ATX4 functional studies beyond Western blotting?
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RNA-seq integration: Identify ATX4-regulated genes by comparing transcriptomes of WT and atx4 mutants under ABA stress .
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CUT&Tag: Achieve higher-resolution mapping of ATX4 binding sites compared to traditional ChIP-seq.
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Structural modeling: Use databases like AACDB to analyze ATX4’s epitope-paratope interface and predict antibody-antigen interactions .
How do I design experiments to map ATX4 antibody epitopes?
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Peptide arrays: Synthesize overlapping 15-mer peptides spanning ATX4’s C-terminal domain (critical for H3K4 methylation) .
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Competitive ELISA: Compare binding affinity of ATX4 antibody with/without pre-incubation with recombinant ATX4 fragments .
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CRISPR-Cas9 truncations: Generate Arabidopsis lines with deletions in ATX4’s catalytic SET domain to confirm antibody targeting .
What bioinformatics tools are essential for analyzing ATX4-associated genomic data?
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AACDB: Query antigen-antibody structural data to refine epitope predictions .
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MEME Suite: Identify conserved motifs in ATX4-bound genomic regions from ChIP-seq data .
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STRING DB: Construct protein interaction networks linking ATX4 to ABA signaling components (e.g., ABI3, ABI5) .
How can I optimize ATX4 antibody storage for long-term stability?
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Lyophilization: Reconstitute antibodies in 50% glycerol/PBS to prevent freeze-thaw degradation .
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Validation schedule: Re-test titer and specificity every 6 months using standardized WT/atx4 mutant lysates .
What strategies mitigate off-target effects in ATX4 knockdown experiments?
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Complementation lines: Express RNAi-resistant ATX4 transgenes in atx4 mutants to confirm phenotype rescue .
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Single-cell sequencing: Profile cell-type-specific ATX4 effects to distinguish direct vs. indirect transcriptional changes.
Key Findings from Recent Studies
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ATX4 directly trimethylates H3K4 at AHG3, modulating ABA-responsive gene silencing .
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Antibodies targeting ATXβ (a classical ATX isoform) show no cross-reactivity with novel isoforms (ATXδ/ε) in serum assays .
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Structural databases like AACDB reveal conserved paratope residues in ATX4 antibodies critical for epitope recognition .
