GA20OX5 Antibody
Description
GA20ox5: Biochemical and Functional Overview
GA20ox5 is one of five GA20ox genes in Arabidopsis thaliana and other plants, encoding enzymes that catalyze sequential oxidation steps in GA biosynthesis (e.g., converting GA₁₂ to GA₉) . Key findings include:
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Catalytic Activity: GA20ox5 exhibits partial activity, stopping at intermediate GA₂₄ and GA₂₅, unlike GA20ox1–GA20ox4, which fully convert GA₁₂ to GA₉ .
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Developmental Roles: GA20ox5 has minor contributions to plant growth and fertility compared to GA20ox1–GA20ox3, which are critical for stem elongation and floral development .
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Expression Patterns: GA20ox5 transcripts are less abundant in vegetative tissues and are not strongly tied to GA-dependent growth stages .
Antibodies for GA20ox Proteins: Current Status
While antibodies for GA20ox enzymes exist, none are explicitly reported for GA20ox5:
| Antibody ID | Target | Reactivity | Source |
|---|---|---|---|
| AS09 483 | GA20ox (general) | Arabidopsis thaliana | |
| AS11 1631 | GA20ox (general) | Arabidopsis thaliana | |
| AS14 2800 | GID1c (GA receptor) | Arabidopsis thaliana |
Key Observations:
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Existing antibodies target GA20ox broadly or GA receptors (e.g., GID1c), not GA20ox5 specifically .
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No studies in the provided literature validate GA20ox5-specific antibody binding or application.
Research Gaps and Challenges
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Lack of GA20ox5-Specific Tools:
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Alternative Detection Methods:
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Future Directions:
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Epitope Mapping: GA20ox5’s unique sequence features (e.g., regions lacking full catalytic domains) could guide antibody development.
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Tissue-Specific Studies: Antibodies against GA20ox5 may aid in dissecting its role in non-canonical GA pathways (e.g., stress responses).
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Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
Q&A
Here’s a structured collection of FAQs tailored for academic researchers investigating GA20OX5 antibodies, incorporating methodological guidance and data-driven insights from published research:
What experimental controls are critical when analyzing GA20OX5 protein levels across developmental stages?
Advanced considerations:
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Internal controls: Normalize GA20OX5 signals to housekeeping proteins (e.g., actin) and include biological replicates (n ≥ 5) to account for tissue heterogeneity .
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Negative controls: Use tissues from GA20ox5 loss-of-function mutants or RNAi lines.
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Time-course sampling: Collect samples at consistent circadian timepoints to minimize diurnal gibberellin fluctuation effects .
How can I resolve discrepancies between GA20OX5 mRNA and protein abundance data?
Troubleshooting approach:
What are optimal fixation methods for GA20OX5 immunocytochemistry in meristematic tissues?
Protocol optimization:
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Test paraformaldehyde (4%) vs. methanol:acetic acid (3:1) fixation. Methanol-based fixes better preserve small molecule antigens like GA intermediates .
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Include pectinase pretreatment (0.1% w/v, 30 min) to improve antibody penetration in lignified tissues .
How to design a study linking GA20OX5 activity to phenotypic outcomes in transgenic plants?
Multidisciplinary design:
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Genetic: Compare GA20OX5 overexpression lines with CRISPR-Cas9 knockouts .
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Hormonal: Quantify GA4 (the primary bioactive product of GA20OX5) via LC-MS/MS at floral transition stages .
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Phenotypic: Measure internode elongation rates and flowering time under short-day conditions .
What orthogonal methods confirm GA20OX5 antibody reliability in mutant screens?
Validation pipeline:
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Step 1: Antibody-based ELISA vs. enzymatic activity assays (measuring GA20-oxidation rates in vitro).
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Step 2: Spatial correlation with GA20OX5 promoter-GUS reporter lines .
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Step 3: Single-cell RNA sequencing to verify protein-mRNA concordance in shoot apical meristems .
How does GA20OX5 antibody performance vary across plant species?
Cross-species reactivity data:
What statistical approaches address high variability in GA20OX5 western blot quantitation?
Advanced analysis framework:
