Customize GA2OX2 Antibody

Description

Introduction to GA2OX2 Antibody

Gibberellin 2-beta-Dioxygenase 2 (GA2OX2) is a key enzyme in the gibberellin (GA) biosynthesis pathway, responsible for inactivating bioactive GAs (e.g., GA₄ and GA₁) through 2-oxidation. The GA2OX2 antibody is a research tool designed to detect and quantify this enzyme, enabling studies on its role in plant development, hormone regulation, and stress responses. Below is a detailed analysis of its characteristics, applications, and research findings.

GA2OX2 Antibody Characteristics

The GA2OX2 antibody is typically generated using recombinant GA2OX2 protein as an antigen. Key properties include:

Property	Detail	Source
Host Organism	Pisum sativum (garden pea) or Arabidopsis thaliana (model plant)
Expression System	Yeast (Saccharomyces cerevisiae)
Tag	His tag (for purification and detection)
Purity	>90% (via chromatographic techniques)
Concentration	0.2–2 mg/mL (in Tris-based buffer with 50% glycerol)
Storage	Lyophilized; store at -20°C (or -80°C for extended periods)

Gene Function and Biological Role

GA2OX2 regulates GA levels by converting active GAs to inactive forms, influencing critical plant processes:

Process	Mechanism	Evidence
Seed Germination	Elevated GA2OX2 expression reduces bioactive GAs, delaying germination.	GA2ox2 mutants show increased GA levels and faster germination .
Root Development	GA2OX2 inactivation promotes root cell elongation by maintaining low GA.	HDT1/2 mutants (repressors of GA2ox2) show reduced root cell expansion .
Stress Responses	GA2OX2 activity modulates GA turnover under stress conditions.	Overexpression of GA2ox2 alters GA sensitivity in abiotic stress .

Research Applications of GA2OX2 Antibody

The antibody is utilized in studies to:

Quantify GA2OX2 Protein Levels:
- ELISA: Detects GA2OX2 in plant extracts to correlate enzyme abundance with developmental stages or stress conditions .
- Western Blot: Validates protein expression in transgenic plants or mutants (e.g., hdt1,2i mutants with upregulated GA2ox2) .
Characterize Gene Regulation:
- ChIP-seq: Identifies transcription factors (e.g., GAF1, HDT1/2) binding to the GA2ox2 promoter to regulate its expression .
- Immunoprecipitation: Maps protein interactions (e.g., DELLA-GAF1 complex binding to GA2ox2) .
Study Phytohormone Crosstalk:
- Co-immunoprecipitation: Investigates GA2OX2 interactions with GA receptors (e.g., GID1) or other hormones (e.g., auxins, cytokinins) .

Regulation of GA2OX2 Expression

HDT1/2 (Histone Deacetylases): Repress GA2ox2 transcription by reducing histone acetylation at its promoter, delaying root cell expansion .
DELLA Proteins: Interact with GAF1 to bind GA2ox2 promoters, enabling GA feedback regulation of gene expression .

Functional Impact in Plants

Experimental Model	Phenotype	Mechanism	Reference
hdt1,2i Mutants	Reduced root meristem cell number	Upregulated GA2ox2 → lower GA levels → premature cell expansion
GA2ox2 Overexpressors	Dwarfism, delayed flowering	Excessive GA inactivation → growth inhibition

Technological Advancements

Recombinant Protein Production: Yeast systems are preferred for cost-effective GA2OX2 production, enabling large-scale antibody generation .

Product Specs

Buffer

**Preservative:** 0.03% Proclin 300
**Constituents:** 50% Glycerol, 0.01M PBS, pH 7.4

Form

Liquid

Lead Time

Made-to-order (14-16 weeks)

Synonyms

GA2OX2 antibody; At1g30040 antibody; T1P2.6Gibberellin 2-beta-dioxygenase 2 antibody; EC 1.14.11.13 antibody; GA 2-oxidase 2 antibody; Gibberellin 2-beta-hydroxylase 2 antibody; Gibberellin 2-oxidase 2 antibody

Target Names

GA2OX2

Uniprot No.

Q9XFR9

Target Background

Function

This antibody catalyzes the 2-beta-hydroxylation of several biologically active gibberellins, resulting in the homeostatic regulation of their endogenous levels. Gibberellin (GA) catabolism is crucial for plant development. This antibody converts GA9/GA20 to GA51/GA29 and GA4/GA1 to GA34/GA8.

Database Links

KEGG: ath:AT1G30040

STRING: 3702.AT1G30040.1

UniGene: At.412

Protein Families

Iron/ascorbate-dependent oxidoreductase family, GA2OX subfamily

Tissue Specificity

Preferentially expressed in flowers, siliques, and upper stems. Expressed in cotyledons, at the base of the shoot apical meristem and developing leaf primordia.

Q&A

FAQs for GA2OX2 Antibody in Academic Research

Advanced Research Questions

How to design experiments investigating GA2OX2’s role in gibberellin (GA) homeostasis under nitrogen limitation?

Strategy:
- Genetic: Compare GA metabolite profiles (via LC-MS) in WT vs. GA2OX2-overexpressing lines under low-N conditions .
- Transcriptional: Use qRT-PCR to analyze feedback regulation of GA20ox and GA3ox genes .
Key Consideration: Account for tissue-specific GA2OX2 expression (e.g., roots vs. shoots) using compartmentalized sampling .

How to resolve contradictory subcellular localization data for GA2OX2?

Methodological Framework:
- Hypothesis: GA2OX2 may localize differently depending on developmental stage or stress.
- Approach:
  - Perform immunofluorescence with high-resolution confocal microscopy.
  - Use subcellular fractionation (e.g., cytosolic vs. nuclear extracts) followed by Western blot .
- Controls: Co-stain with organelle-specific markers (e.g., GFP-tagged nuclear/cytosolic proteins).

How to address cross-reactivity with GA2OX isoforms in non-model species?

Solution:
- Generate phylogenetic alignment of GA2OX sequences from target species (e.g., using UniProt Q9XHM5 as reference).
- Synthesize peptide antigens unique to GA2OX2 for custom antibody production .
Data Table:

Species	GA2OX2 Homology (%)	Cross-Reactivity Risk
Arabidopsis	100% (Reference)	Low
Oryza sativa	78%	High (test with knockout lines)

Methodological Integration

How to combine GA2OX2 antibody studies with transcriptomic data?

Workflow:
- Perform RNA-seq to identify co-expressed genes (e.g., CCA1, GLK1) under GA2OX2 perturbation .
- Validate protein-level changes via Western blot and correlate with mRNA abundance.
Pitfall: Post-translational modifications (e.g., phosphorylation) may decouple mRNA-protein correlations.

What controls are essential for GA2OX2 functional studies in mutant backgrounds?

Required Controls:
- Biological: At least three independent transgenic lines.
- Technical: Include empty vector controls and rescue lines (e.g., GA2OX2 complementation) .
- Phenotypic: Monitor plant height and seed germination rates, as GA2OX2 inactivation reduces bioactive GA levels .

Data Interpretation Challenges

How to distinguish GA2OX2-specific effects from broader GA pathway perturbations?

Strategy: Use combinatorial mutants (e.g., ga2ox2 ga3ox1) and compare phenotypes to single mutants. Measure GA intermediates (e.g., GA₄, GA₃₄) to pinpoint metabolic bottlenecks .

Why might GA2OX2 antibody signals vary across tissue types?

Explanation: GA2OX2 expression is regulated by N-status and circadian rhythms (via CCA1) . Standardize sampling to a consistent time-of-day and nutrient condition.

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GA2OX2 Antibody