At3g09930 Antibody
Description
Functional Role of AT3G09930
The AT3G09930 gene encodes an enzyme critical for:
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Lipid remodeling: Hydrolysis and synthesis of acyl groups in glycerolipids.
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Stress response: Modulation of lipid membranes under environmental stressors.
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Seed development: Involvement in endosperm localization, as demonstrated by immunolocalization studies .
The antibody specifically recognizes epitopes on AT3G09930 and cross-reacts with related proteins MIPS1, MIPS2, and MIPS3, which are implicated in myo-inositol biosynthesis .
Research Applications
The At3g09930 antibody has been utilized in diverse experimental workflows:
Key Research Findings
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Endosperm Localization: The antibody revealed AT3G09930’s presence in endosperm cells, linking it to lipid droplet formation during seed maturation .
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Stress Adaptation: Studies using this antibody showed upregulated AT3G09930 expression during drought and pathogen exposure, indicating its role in membrane lipid restructuring .
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Interaction Network: Co-immunoprecipitation experiments identified interactions with phospholipase Dζ2 and lipid-transfer proteins, highlighting its centrality in lipid signaling pathways .
Technical Considerations
Product Specs
Composition: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Target Background
Q&A
The At3g09930 antibody is a research tool primarily used in plant molecular biology studies, particularly in Arabidopsis thaliana research. Below is a structured FAQ addressing both foundational and advanced research considerations, incorporating methodological guidance and hypothetical data tables based on typical antibody validation paradigms.
Advanced Research Applications
4. Resolving contradictory results between Western blot and qPCR data
Implement a triangulation approach:
| Step | Method | Critical Parameters |
|---|---|---|
| 1 | Parallel protein/RNA extraction | RNase-free conditions |
| 2 | Spike-in control | Arabidopsis ubiquitin conjugase |
| 3 | Epitope mapping | Compare antibody vs predicted antigenic sites |
Optimizing ChIP-seq protocols for low-abundance targets
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Use crosslinking enhancement: 2% formaldehyde + 5mM EGS (ethylene glycol bis(succinimidyl succinate))
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Sequential chromatin shearing: 15 cycles (30s ON/30s OFF) in Bioruptor®
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Validate with input normalization and IgG controls
6. Quantitative analysis limitations in developmental stage comparisons
Develop a normalization matrix:
| Tissue Type | Correction Factor | Reference Protein |
|---|---|---|
| Meristem | 1.8x | Histone H3 |
| Senescent | 0.6x | Actin |
Methodological Considerations
Troubleshooting nonspecific bands in Western blots
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Implement 2D gel electrophoresis prior to transfer
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Test alternative blocking buffers:
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5% non-fat milk (standard)
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3% BSA + 1% fish gelatin (high specificity)
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Commercial blocking agents (e.g., SEA BLOCK™)
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8. Cross-reactivity assessment in non-Arabidopsis species
Conduct phylogenetic ELISA:
| Species | % Identity | Signal Intensity |
|---|---|---|
| Brassica napus | 89% | +++ |
| Oryza sativa | 67% | + |
| Zea mays | 58% | - |
