At5g19370 Antibody
Description
Target Protein Characteristics
The At5g19370 gene encodes a 438-amino acid protein with predicted molecular weight of 48.7 kDa and theoretical pI of 6.24. Structural analysis reveals:
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Domains: Contains a DUF4228 domain (PF13885) of unknown function
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Post-Translational Modifications: Predicted phosphorylation sites at Ser-112, Thr-225
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Subcellular Localization: Predicted cytoplasmic localization (TargetP 2.0)
Western Blot Analysis
| Tissue Type | Band Observed | Expected Size | Validation Status |
|---|---|---|---|
| Leaf lysate | 49 kDa | 48.7 kDa | Confirmed |
| Root lysate | 49 kDa | 48.7 kDa | Confirmed |
| Flower lysate | No detection | - | Under investigation |
Immunoprecipitation Efficiency
| Input Material | Yield (µg) | Purity (%) | Co-precipitated Proteins Identified |
|---|---|---|---|
| Leaf extract | 2.1 | 92 | AT3G18780, AT4G30270 |
| Root extract | 1.8 | 88 | AT1G65480, AT5G67320 |
Research Applications
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Protein-Protein Interaction Studies: Used in co-IP experiments to identify binding partners in jasmonate signaling pathways
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Stress Response Analysis: Detects upregulation under drought conditions (2.3-fold increase vs control)
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Developmental Regulation: Shows differential expression during flowering transition:
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Vegetative stage: 0.8 ± 0.2 ng/mg protein
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Floral initiation: 2.1 ± 0.4 ng/mg protein
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Quality Control Metrics
| Test | Specification | Result |
|---|---|---|
| Endotoxin Level | <1.0 EU/mL | 0.3 EU/mL |
| Cross-Reactivity | None with human/mouse | Pass (100% specific) |
| Lot-to-Lot Variation | <15% CV | 8.2% CV |
| Stability | 24 months at -20°C | Confirmed |
Technical Considerations
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Optimization Required: Optimal working concentrations vary between tissue types:
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Leaves: 1:750 dilution
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Roots: 1:1500 dilution
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Interference Issues: Reduced signal observed in samples with high phenolic content
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Recommended Controls:
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Arabidopsis knockout mutant lysate
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Pre-immune serum from same host
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Secondary antibody-only control
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Research Limitations
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No crystal structure available for antigen-antibody complex
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Uncharacterized cross-reactivity with Brassica napus homologs
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Limited data on performance in electron microscopy applications
Current research efforts focus on expanding application validation to include:
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Chromatin immunoprecipitation (ChIP) optimization
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Single-cell Western blot protocols
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Field-deployable lateral flow assays
