At2g19050 Antibody
Description
1. Overview of At2g19050 Antibody
The At2g19050 antibody is a monoclonal or polyclonal immunoglobulin designed to specifically target the Arabidopsis thaliana GDSL esterase/lipase enzyme encoded by the At2g19050 gene. This antibody is primarily used in plant biology research to study the expression, localization, and functional roles of the At2g19050 protein, which is implicated in lipid metabolism, cell wall modification, and stress responses .
3. Target Protein: Functional Insights
The At2g19050 gene encodes a GDSL esterase/lipase, a class of enzymes involved in:
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Regulation of cuticular lipid deposition in plant epidermal cells.
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Response to abiotic stressors (e.g., drought, pathogen attack) .
Structural studies suggest GDSL enzymes have a conserved catalytic triad (Ser, Asp, His) critical for enzymatic activity, though direct structural data for At2g19050 remains limited .
4. Research Applications
Protein Detection and Quantification
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Western Blot: Used to detect At2g19050 protein in Arabidopsis lysates, with a typical band size of ~35–40 kDa (predicted molecular weight) .
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ELISA: Quantifies protein levels in plant tissue extracts under varying experimental conditions .
Subcellular Localization
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Antibody-based immunofluorescence can map At2g19050 distribution in plant tissues, particularly in epidermal cells and vascular bundles .
Functional Studies
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Investigates gene knockout or overexpression phenotypes (e.g., lipid accumulation, stress tolerance) .
5. Validation and Performance Considerations
While commercial antibodies like At2g19050 (CSB-PA524030XA01DOA) are widely used, independent validation is critical. Key steps include:
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Specificity Testing: Confirm absence of cross-reactivity with unrelated Arabidopsis proteins (e.g., via knockout mutant controls) .
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Batch Consistency: Ensure reproducibility across antibody lots .
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Application Optimization: Titrate antibody concentrations for WB/ELISA to minimize background noise .
7. Limitations and Challenges
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Species Specificity: Reactivity is confined to Arabidopsis thaliana; cross-species applications require validation .
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Antibody Validation: As highlighted in studies on angiotensin II receptor antibodies , commercially available antibodies may exhibit off-target binding without rigorous testing.
8. Future Directions
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Structural Studies: Cryo-EM or X-ray crystallography to resolve At2g19050’s 3D structure.
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Pathway Analysis: Integration with omics datasets to elucidate metabolic networks involving GDSL enzymes.
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Agricultural Biotechnology: Engineering At2g19050 overexpression in crops for enhanced stress resilience .
Product Specs
Target Background
KEGG: ath:AT2G19050
STRING: 3702.AT2G19050.1
Q&A
Here’s a structured collection of FAQs tailored for researchers working with the At2g19050 Antibody in academic contexts, based on protein function, validation methodologies, and experimental design principles:
How can I validate the specificity of the At2g19050 antibody in plant tissue extracts?
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Method: Combine Western blotting with Arabidopsis thaliana knockout mutants (e.g., T-DNA insertion lines) as negative controls. Include recombinant At2g19050 protein (NP_179495.1) as a positive control .
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Troubleshooting: If nonspecific bands appear, pre-absorb the antibody with recombinant protein or use tissue from overexpression lines to confirm signal saturation.
What experimental applications are most suitable for this antibody?
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Primary uses:
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Limitations: Avoid flow cytometry for intracellular targets unless fixation/permeabilization protocols are optimized.
How should I quantify At2g19050 protein levels across developmental stages?
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Workflow:
How do I resolve contradictions in subcellular localization data?
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Scenario: Antibody signals in cytoplasmic compartments vs. extracellular regions .
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Approach:
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Perform subcellular fractionation followed by Western blotting.
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Validate with fluorescent protein-tagged At2g19050 constructs (e.g., GFP fusions).
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What controls are critical for lipid interaction assays using this antibody?
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Experimental design:
How can I differentiate between isoforms or post-translationally modified forms?
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Strategy:
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Use 2D gel electrophoresis + Western blotting to separate charge variants.
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Treat samples with phosphatases (e.g., λ-PPase) or glycosidases (e.g., PNGase F) before analysis.
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How to address inconsistent activity correlations in lipase assays?
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Potential factors:
Factor Diagnostic Test Protein stability Cycloheximide chase + antibody-based degradation assay Redox state DTT/TCEP treatment during extraction Phosphorylation Phos-tag SDS-PAGE
What orthogonal methods validate antibody-dependent phenotypes?
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Multi-omics integration:
