At4g16820 Antibody
Description
Introduction to At4g16820 Antibody
The At4g16820 Antibody is a molecular tool designed to detect the protein encoded by the Arabidopsis thaliana gene At4g16820, a phospholipase A I beta 2 (PLA-Iβ2) enzyme. This gene, also known as DALL1 (Defective in Anther Dehiscence 1-like Lipase 1), encodes a lipase involved in lipid metabolism, stress responses, and jasmonic acid (JA) biosynthesis. The antibody facilitates research into its role in membrane lipid processing, plant defense, and lipid-based signaling pathways .
Functional Role of At4g16820 Protein
The At4g16820 protein belongs to the alpha/beta-Hydrolases superfamily and hydrolyzes phosphatidylcholine, glycolipids (e.g., MGDG, DGDG), and triacylglycerols (TAGs) . Key functions include:
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Lipid Metabolism: Catalyzing the release of free fatty acids (FFAs) from membrane lipids, which are precursors for oxylipin signaling molecules like JA .
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Stress Responses: Participating in wound-induced JA biosynthesis and salt stress adaptation .
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Lipid Signaling: Facilitating lipid transport in phloem exudates, potentially enabling long-distance signaling .
Detection in Stress and Developmental Studies
The antibody is used to immunodetect At4g16820 protein levels in tissues under stress (e.g., wounding, salinity) or during developmental processes. For example:
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Western Blotting: Validates protein expression in mutants or transgenic plants. In PDEX:AtDAD1-Myc lines, Myc-tagged DAD1-like proteins are analyzed to study lipid remodeling .
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Immunolocalization: Identifies subcellular localization (e.g., chloroplasts, membranes) .
Role in Lipid Remodeling Pathways
Studies using antibodies to detect related lipases (e.g., DAD1) reveal:
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TAG Accumulation: Overexpression of DAD1-like enzymes redirects fatty acids into TAGs via PDAT1/DGAT1 pathways .
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Lipid Profiling: MGDG (18:3/18:3) and DGDG (36:6) levels decrease post-treatment, while LysoPC species accumulate .
Gene Expression and Lipid Changes
Protein Interactions and Regulation
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COI1 Dependency: In dgd1 mutants, At4g16820 activity is influenced by COI1-mediated JA feedback loops, altering LOX/AOC enzyme levels .
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Pathway Crosstalk: Contributes to JA biosynthesis alongside PLA-Iγ3 and LOX enzymes .
Technical Considerations for Antibody Use
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Specificity: The antibody may cross-react with related lipases (e.g., DAD1), requiring validation via peptide blocking or mutant controls .
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Tissue Optimization: Highest expression in leaves and flowers, with induced levels in roots under stress .
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Commercial Availability: Limited to custom synthesis, as no commercial antibody is explicitly reported in literature.
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
KEGG: ath:AT4G16820
STRING: 3702.AT4G16820.1
Q&A
Here’s a structured FAQ for researchers investigating the At4g16820 Antibody in Arabidopsis thaliana studies, incorporating methodological guidance and data-driven insights:
What are the key applications of At4g16820 Antibody in studying salt stress tolerance?
Experimental Design:
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Use immunolocalization to track subcellular redistribution of At4g16820 under high-salinity conditions (e.g., 150 mM NaCl) .
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Pair with qRT-PCR to correlate protein abundance with transcriptional changes in related pathways (e.g., SOS1, SOS3) .
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Integrate with ChIP-seq to identify DNA-binding targets under stress .
Advanced Insight:
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At4g16820 may interact with HDAC inhibitors (e.g., Ky-9/Ky-72) but not SOS pathway components, suggesting alternative regulatory mechanisms .
How to resolve contradictory data between antibody detection and transcriptomic profiles?
Troubleshooting Framework:
Case Example:
In HDAC inhibitor studies, At4g16820 protein increased despite unchanged mRNA, implicating post-translational regulation .
What controls are essential for co-immunoprecipitation assays using this antibody?
Required Controls:
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Isotype-matched IgG to exclude nonspecific binding
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Bait-only lysate (transfected empty vector)
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Pre-immune serum from the same host species
Data Quality Metric:
How to optimize At4g16820 Antibody for multiplexed imaging workflows?
Protocol Optimization:
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Cross-adsorption: Pre-incubate with A. thaliana protein extracts to remove species-specific cross-reactivity .
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Signal Amplification: Combine tyramide-based amplification with ≤2 other fluorophores (e.g., Alexa 488/647).
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Validation: Compare staining patterns across developmental stages using CLSM with Airyscan detection .
What computational tools integrate At4g16820 Antibody data with transcriptional networks?
Advanced Workflow:
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Map protein localization data to AraPPIS (Arabidopsis Protein-Protein Interaction Server)
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Overlay with RNA-seq clusters using STEM (Short Time-series Expression Miner)
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Predict regulatory hierarchies via SDREM (Stochastic Discrete Regulation Modeling)
Key Finding:
At4g16820 clusters with pathogen-response modules but not ion homeostasis pathways, aligning with its absence in SOS1/3 regulation .
