At5g62930 Antibody
Description
Definition and Biological Context
At5g62930 Antibody is a specialized immunological reagent targeting the protein encoded by the At5g62930 gene in Arabidopsis thaliana (thale cress). This gene encodes an SGNH hydrolase-type esterase, a class of enzymes involved in lipid metabolism and plant defense mechanisms . The antibody is designed to detect and bind specifically to this protein, enabling researchers to study its expression, localization, and functional roles in plant biology.
Role of At5g62930 in Plant Biology
The At5g62930 protein belongs to the SGNH hydrolase superfamily, which is characterized by conserved serine, glycine, asparagine, and histidine residues critical for catalytic activity . These enzymes are implicated in:
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Lipid metabolism: Hydrolysis of ester bonds in fatty acid derivatives .
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Stress responses: Modulating plant defense against pathogens .
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Cell wall remodeling: Facilitating structural changes during growth .
Antibody Validation and Cross-Reactivity
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Specificity: The antibody binds selectively to the At5g62930 epitope, as demonstrated in Western blot analyses of Arabidopsis extracts .
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Cross-Reactivity: No significant cross-reactivity with homologous proteins (e.g., At5g03610 or At4g28780) has been reported .
Protein Localization Studies
The At5g62930 Antibody has been employed to map the subcellular distribution of the protein in Arabidopsis tissues. Immunohistochemical assays reveal strong signals in vascular tissues and root meristems, suggesting roles in nutrient transport and developmental regulation .
Transcriptional Regulation Analysis
Promoter-binding studies using transgenic Arabidopsis lines indicate that At5g62930 expression is influenced by cytokinin signaling pathways. The antibody aids in tracking protein levels under hormonal treatments .
Comparative Genomics
Comparative analyses in pecan (Carya illinoinensis) identified orthologs of At5g62930 (e.g., CilGDSL27.35), highlighting conserved functions across plant species .
Stress Adaptation
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Cold Tolerance: Knockout mutants of At5g62930 exhibit delayed senescence under cold stress, implicating the protein in stress resilience .
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Pathogen Response: SGNH hydrolases like At5g62930 are upregulated during fungal infections, suggesting a role in hydrolyzing pathogen-derived lipids .
Metabolic Pathways
Enzymatic assays using purified At5g62930 protein demonstrate activity against acetylated xyloglucans, linking it to cell wall modification .
Limitations and Future Directions
Product Specs
Constituents: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Q&A
FAQs for Researchers on At5g62930 Antibody
Advanced Research Questions
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How to resolve contradictions between antibody performance in Western blot vs. flow cytometry?
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Methodology:
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Hypothesis: Post-translational modifications (e.g., glycosylation) may alter epitope accessibility. Test deglycosylation (e.g., PNGase F treatment) in Western blot and compare with flow cytometry results .
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Experimental design:
Assay Type Glycosylation Status Antibody Binding (Relative Intensity) Western Native 1.0 Western Deglycosylated 0.2 Flow Cytometry Native 0.8
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What strategies improve antibody-dependent cellular cytotoxicity (ADCC) for therapeutic At5g62390-targeting antibodies?
Troubleshooting & Data Analysis
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How to address nonspecific binding in immunohistochemistry (IHC) with At5g62390 antibodies?
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What computational tools predict At5g62390-antibody binding dynamics?
Experimental Design
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How to standardize At5g62390 antibody dilution across assays?
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How to validate antibody functionality in 3D cell culture models?
Key Considerations:
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Cross-reactivity: Always pair antibody validation with orthogonal techniques (e.g., CRISPR validation + mass spectrometry) .
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Assay context: Antibody performance may vary between denatured (Western) vs. native (flow cytometry/IHC) conditions .
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Therapeutic potential: Prioritize antibodies with dual mechanisms (e.g., ADCC enhancement and receptor blockade) .
