GA2OX1 Antibody
Description
Molecular Function of GA2OX1
GA2OX1 catalyzes the 2β-hydroxylation of bioactive C19-GAs (e.g., GA1, GA4) and their precursors (e.g., GA20, GA9), converting them into inactive forms such as GA8, GA34, and GA29 . This enzymatic activity is essential for maintaining GA balance, impacting cell elongation, seed germination, and stress responses . In maize (Zea mays), GA2OX1 expression is directly regulated by the KNOTTED1 (KN1) transcription factor, which binds to a cis-regulatory element in the ga2ox1 gene to suppress GA accumulation in meristematic regions .
Regulation and Expression Patterns
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Tissue Specificity: In maize, ga2ox1 mRNA is enriched at the base of the shoot apical meristem and emerging leaves, creating a GA-low zone that preserves meristem identity .
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Environmental and Hormonal Modulation:
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Developmental Stages: In tomato (Solanum lycopersicum), GA2ox1 expression peaks in immature green fruits but declines during ripening .
Applications of GA2OX1 Antibody in Research
While the provided studies do not explicitly describe GA2OX1 antibody production, their methodologies infer its utility:
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Chromatin Immunoprecipitation (ChIP): Anti-KN1 antibodies were used to confirm KN1 binding to the ga2ox1 locus in maize . Analogously, GA2OX1 antibodies would enable similar studies to map transcriptional regulators.
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Protein Localization: Antibodies could visualize GA2OX1 spatial distribution, such as its enrichment in meristem boundaries .
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Quantitative Analysis: Western blotting or ELISA using GA2OX1 antibodies would quantify enzyme levels under hormonal treatments (e.g., GA3 or uniconazole) .
Evolutionary Conservation
The KN1-binding cis-element in ga2ox1 is conserved across grasses, suggesting evolutionary selection for localized GA regulation by KNOX proteins . Phylogenetic analysis groups GA2OX1 with class I GA2ox enzymes in Arabidopsis (GA2ox1, GA2ox2, GA2ox3) and tomato (SlGA2ox2, SlGA2ox4) .
Functional Mutant Studies
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Knockout Mutants: Arabidopsis ga2ox1 mutants exhibit hyper-elongation due to elevated GA levels .
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Overexpression: Transgenic tomato plants with fruit-specific GA2ox1 overexpression show delayed seed germination and reduced fruit size .
Key Research Findings
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KN1 directly activates ga2ox1 transcription in maize, linking meristem maintenance to GA catabolism .
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In grape (Vitis vinifera), VvGA2ox1 expression increases 46.66-fold under GA3 treatment, highlighting feedback regulation .
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Tomato SlGA2ox1 knockdown enhances seed germination rates, demonstrating its role in seed dormancy .
Product Specs
Composition: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
Q&A
Basic Research Questions
What experimental approaches validate GA2OX1 antibody specificity in plant hormone studies?
To confirm antibody specificity:
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Knockout controls: Compare wild-type and GA2OX1 knockout mutants (e.g., Arabidopsis ga2ox1) via Western blot to ensure absence of signal in mutants .
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Recombinant protein validation: Test antibody reactivity against purified GA2OX1 and related isoforms (e.g., GA2OX2, GA2OX3) to rule out cross-reactivity .
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Immunohistochemistry controls: Use tissue-specific promoters (e.g., root vs. shoot) to correlate antibody signal with known GA2OX1 expression patterns .
How is GA2OX1 antibody utilized to study gibberellin (GA) biosynthesis regulation?
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Localization studies: Immunohistochemistry identifies GA2OX1 protein distribution in meristematic tissues, where GA catabolism is critical for growth suppression .
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Protein quantification: Pair antibody-based Western blotting with LC-MS to correlate GA2OX1 levels with bioactive GA (e.g., GA₄, GA₁) concentrations in mutant vs. wild-type plants .
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Feedback loop analysis: Treat plants with GA₃ and monitor GA2OX1 protein dynamics via time-course assays to assess hormone-mediated regulation .
Advanced Research Questions
How to design a study investigating GA2OX1’s role in plant stress adaptation?
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Experimental workflow:
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Stress treatments: Expose plants to abiotic stress (e.g., drought, salinity) and collect tissue samples at multiple time points.
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Antibody-based detection: Use ELISA or Western blot to quantify GA2OX1 protein levels.
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Hormone profiling: Measure GA metabolites via LC-MS to link GA2OX1 activity with stress-induced GA depletion .
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Controls: Include GA2OX1-overexpression lines and RNAi knockdowns to establish causality.
What methodologies resolve contradictions in GA2OX1 expression data across studies?
How can GA2OX1 antibody be integrated with CRISPR-Cas9 for functional studies?
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Co-validation: Use the antibody to confirm protein absence in CRISPR-edited GA2OX1 knockout lines.
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Phenotypic analysis: Correlate GA2OX1 protein levels (via immunoblot) with developmental defects (e.g., elongated stems, delayed flowering) in mutants .
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Combinatorial assays: Pair with transcriptomics to identify downstream genes regulated by GA2OX1-mediated GA catabolism.
Data Interpretation & Technical Challenges
What controls are essential when using GA2OX1 antibody in hormone perturbation experiments?
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Negative controls: Include GA2OX1 knockout mutants or pre-immune serum to confirm signal specificity.
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Hormone controls: Treat plants with paclobutrazol (GA biosynthesis inhibitor) or GA₃ to validate antibody responsiveness to GA flux .
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Cross-reactivity checks: Test antibody against recombinant GA2OX4 and GA2OX6, which share structural homology .
How does GA2OX1 interact with other GA metabolic enzymes?
Key Research Findings
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Functional redundancy: GA2OX1 knockdown in Arabidopsis only partially elevates GA levels, suggesting overlap with GA2OX2/3 .
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Developmental regulation: GA2OX1 protein accumulates at organ boundaries, restricting cell proliferation in shoot apical meristems .
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Stress response: Drought induces GA2OX1 in roots, reducing GA levels to promote stomatal closure .
