At1g31550 Antibody
Description
Introduction to At1g31550 Antibody
The At1g31550 antibody is a specialized immunological tool targeting the protein encoded by the Arabidopsis thaliana gene AT1G31550, which belongs to the GDSL lipase acylhydrolase family. This protein is implicated in lipid metabolism and plant cell wall adaptation under sub-optimal growth conditions. The antibody enables researchers to detect, localize, and quantify the expression of this protein in experimental studies, particularly in plant biology and stress-response research .
Biological Role of At1g31550
The AT1G31550 gene encodes a GDSL-motif lipase/hydrolase, a class of enzymes involved in lipid remodeling and hydrolysis. Key functional attributes include:
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Lipid Metabolism: Catalyzes the breakdown of ester bonds in lipids, influencing membrane composition and signaling .
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Stress Adaptation: Differential accumulation of the AT1G31550 protein under environmental stress suggests a role in Arabidopsis's adaptive responses to sub-optimal growth conditions .
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Cell Wall Dynamics: Linked to modifications in plant cell wall structure, potentially aiding resilience to abiotic stressors .
Research Applications
The At1g31550 antibody is primarily used in:
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Proteomic Studies: Quantifying protein expression levels under varying growth conditions (e.g., via Western blotting).
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Localization Experiments: Identifying tissue-specific or subcellular distribution of the protein (e.g., immunohistochemistry).
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Functional Genomics: Validating gene knockout or overexpression models in Arabidopsis .
Key Experimental Findings
A 2020 study investigating Arabidopsis adaptation to stress conditions revealed critical data about the AT1G31550 protein (Table 1) :
Table 1: Differential Accumulation of AT1G31550 in Arabidopsis Under Stress
| AGI Code | Functional Class | Putative Function | Protein Level | Transcript Level |
|---|---|---|---|---|
| AT1G31550 | LM | Lipase acylhydrolase (GDSL family) | Increased (+) | Decreased (−) |
Key Observations:
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Protein vs. Transcript Dynamics: Increased protein levels (+), despite decreased transcript levels (−), suggest post-transcriptional regulation under stress .
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Functional Class: Categorized under "Lipid Metabolism" (LM), highlighting its role in lipid-associated stress responses .
Challenges in Antibody Specificity
While the At1g31550 antibody is critical for plant research, broader challenges in antibody validation persist. For example, studies on angiotensin II receptor antibodies (e.g., AT1R) have demonstrated that commercial antibodies often lack specificity, detecting off-target proteins or showing inconsistent results across experimental setups . These findings underscore the need for rigorous validation of At1g31550 antibodies using controls such as:
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Knockout plant lines to confirm target specificity.
Future Research Directions
Product Specs
Components: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Target Background
Q&A
Here’s a structured collection of FAQs tailored to academic research scenarios for the At1g31550 Antibody, incorporating methodological guidance, data insights, and technical depth:
What experimental designs are optimal for studying post-translational modifications (PTMs) with At1g31550 Antibody?
Basic Workflow:
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Use phosphate-affinity electrophoresis for phosphorylation studies, coupled with lambda phosphatase treatment to confirm specificity .
Advanced Strategy:
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Pair the antibody with ubiquitination-specific proteasome inhibitors (e.g., MG132) in time-course assays to track degradation dynamics.
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Integrate crosslinking-MS to map interaction sites modified by PTMs .
How to resolve contradictions in quantitative data from At1g31550 Antibody-based assays?
Troubleshooting Framework:
What methods enhance sensitivity in low-abundance target detection?
Basic Approach:
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Use DELFIA® (Dissociation-Enhanced Lanthanide Fluorescent Immunoassay) with europium-labeled secondary antibodies for time-resolved fluorescence detection, improving signal-to-noise ratios .
Advanced Optimization:
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Combine with immuno-PCR for ultrasensitive quantification, leveraging DNA amplification for trace-level detection.
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Validate using serial dilutions of recombinant protein (e.g., 0.1–100 ng/mL) to establish a linear dynamic range .
How to design a multiplex assay involving At1g31550 Antibody?
Methodological Guidance:
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Use spectrally distinct fluorophores (e.g., Cy3, Cy5) for co-staining with other antibodies. Verify emission spectra overlap using tools like FPbase.
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For protein-protein interaction studies, pair with Bimolecular Fluorescence Complementation (BiFC) to visualize real-time interactions .
What controls are critical for CRISPR/Cas9-edited lines in At1g31550 studies?
Essential Controls:
| Control Type | Purpose |
|---|---|
| Wild-type + antibody | Baseline signal |
| Wild-type + no primary antibody | Background assessment |
| Knockout + antibody | Specificity confirmation |
| Heterozygous line | Dosage effect analysis |
Advanced Application:
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Perform single-cell RNA-seq on edited lines to identify compensatory gene networks masking phenotypic effects .
