At4g16220 Antibody
Description
Target Protein: At4g16220
The At4g16220 gene encodes a GDSL-like Lipase/Acylhydrolase superfamily protein (UniProt ID: O23469), which exhibits esterase/lipase activity (EC 3.1.1.-). Key features include:
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Function: Associated with extracellular lipid metabolism, potentially influencing plant cell wall dynamics or stress responses .
Research Applications
The At4g16220 antibody is employed in diverse experimental workflows:
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Western Blot: Used to detect the ~45 kDa At4g16220 protein in Arabidopsis lysates .
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ELISA: Quantifies protein expression levels under varying physiological conditions .
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Functional Studies: Investigates the role of GDSL lipases in plant lipid signaling and stress adaptation .
Technical Considerations
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Specificity: Polyclonal nature ensures recognition of multiple epitopes, enhancing detection sensitivity .
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Cross-Reactivity: No reported cross-reactivity with non-target plant species .
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Limitations: Lack of published validation data in peer-reviewed studies (based on available sources).
Context in Antibody Development
While the At4g16220 antibody itself is not directly discussed in GPCR or therapeutic antibody research, broader studies highlight the importance of polyclonal antibodies in basic science:
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Diversity: Polyclonal antibodies capture conformational and linear epitopes, advantageous for detecting structurally dynamic proteins .
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Engineering: Advances in antibody engineering (e.g., Fc modifications) could enhance utility in plant biochemistry studies, though this remains unexplored for At4g16220 .
Product Specs
Constituents: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Target Background
KEGG: ath:AT4G16220
Q&A
The AT4G16220 antibody is a critical tool in plant molecular biology, particularly for studying Arabidopsis thaliana gene functions. Below is a structured FAQ collection tailored to academic research needs, organized by complexity and methodological focus:
Advanced Research Questions
Resolving contradictory data in AT4G16220 subcellular localization studies
| Issue | Diagnostic Approach | Example Solution |
|---|---|---|
| Nuclear vs. cytoplasmic signals | Fractionation + protease inhibition | Adjust lysis buffer pH to 7.4 |
| Variability across cell types | Cell-type-specific promoters (e.g., WOX5) | Use INTACT isolation |
Optimizing ChIP-seq protocols for AT4G16220-DNA interactions
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Critical parameters:
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Crosslinking time: 15 min vs. 30 min (test via qPCR recovery)
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Sonication efficiency: Aim for 200-500 bp fragments
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Antibody concentration: Titrate from 1:100 to 1:1000
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Integrating AT4G16220 antibody data with CRISPR-Cas9 knockout phenotypes
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Multi-omics validation framework:
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Transcriptomics: RNA-seq of knockout lines
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Proteomics: Antibody-based quantification vs. mass spectrometry
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Phenotypic correlation: Rosette size/root growth metrics
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Methodological Best Practices
Addressing non-specific bands in AT4G16220 Western blots
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Systematic troubleshooting:
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Blocking buffer: Compare 5% BSA vs. non-fat milk
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Secondary antibody: Switch between HRP and AP conjugates
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Membrane type: PVDF vs. nitrocellulose (55 kDa target expected)
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Quantitative analysis of AT4G16220 expression under abiotic stress
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Normalization strategy:
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Internal control: Rubisco large subunit (55 kDa)
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External standard: Recombinant AT4G16220 protein ladder
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Statistical thresholds: ≥2-fold change, p < 0.01 (ANOVA)
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