At3g58980 Antibody
Description
Introduction to At3g58980 Antibody
The At3g58980 Antibody is a rabbit polyclonal antibody designed to target the protein product of the At3g58980 gene in Arabidopsis thaliana (mouse-ear cress). This antibody is primarily used in plant molecular biology research to study the F-box/LRR-repeat protein encoded by the gene, which may play roles in protein degradation or regulatory pathways. The antibody is commercially available from suppliers like MyBioSource and Cusabio, with applications in Western blotting (WB), enzyme-linked immunosorbent assay (ELISA), and immunoprecipitation .
Applications in Research
The At3g58980 Antibody enables targeted detection of its cognate protein in plant tissues. Key applications include:
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Western Blotting (WB):
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Identifies the At3g58980 protein in Arabidopsis lysates.
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Confirms protein expression levels under different experimental conditions (e.g., stress responses).
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ELISA:
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Quantifies At3g58980 protein concentrations in plant extracts or recombinant preparations.
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Immunoprecipitation:
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Isolates At3g58980 for downstream analysis of protein interactions (e.g., F-box protein complexes).
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Limitations:
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Limited data on cross-reactivity with homologous proteins in other species.
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No reported use in immunohistochemistry (IHC) or immunofluorescence (IF) to date .
Comparative Analysis with Other Arabidopsis Antibodies
The At3g58980 Antibody is part of a broader portfolio of antibodies targeting Arabidopsis proteins. Below is a comparison with three representative antibodies from Cusabio :
Key Insight:
The At3g58980 Antibody shares similar technical specifications with other Arabidopsis antibodies but targets a distinct protein family (F-box/LRR-repeat), which may be involved in ubiquitin-mediated protein degradation .
Challenges and Considerations in Antibody Use
While the At3g58980 Antibody is a valuable tool, researchers must address the following:
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Specificity Validation:
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Quantitative Limitations:
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Host-Specific Reactivity:
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Limited to Arabidopsis thaliana; unsuitable for cross-species studies.
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Future Directions and Research Opportunities
The At3g58980 Antibody could advance studies in:
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Protein Degradation Pathways:
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Stress Response Mechanisms:
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Profiling At3g58980 expression under biotic or abiotic stress conditions.
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Antibody Optimization:
Product Specs
Q&A
Here’s a structured collection of FAQs tailored for researchers working with the At3g58980 Antibody in academic settings, integrating technical depth and methodological guidance:
Key Validation Parameters:
What experimental conditions optimize At3g58980 detection in plant tissue sections?
Advanced Considerations:
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Fixation: Use 4% paraformaldehyde for 2 hr to preserve epitopes without over-fixation.
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Antigen retrieval: Optimize citrate buffer pH (6.0–6.5) and heating time (10–15 min) to balance signal intensity and tissue integrity.
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Blocking: 5% BSA + 0.1% Tween-20 reduces non-specific binding in lignified plant tissues .
How to resolve contradictory localization data for At3g58980 across studies?
Analytical Framework:
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Technical variables: Compare fixation methods, antibody dilutions, and imaging platforms used in conflicting studies.
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Biological context: Assess growth conditions (e.g., stress treatments) that may alter protein expression patterns.
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Orthogonal validation: Combine immunofluorescence with in situ hybridization or GFP-tagged transgenic lines .
What statistical approaches are robust for quantifying At3g58980 expression levels?
Methodological Guidance:
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Use linear mixed-effects models to account for batch effects in large-scale experiments.
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Apply background subtraction algorithms (e.g., rolling-ball) to immunoblot data to minimize noise .
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For longitudinal studies, employ ANOVA with repeated measures to track dynamic expression changes.
How to design a CRISPR-based control experiment for At3g58980 functional studies?
Advanced Strategy:
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Generate dual gRNAs targeting exons 2 and 4 of At3g58980 to ensure frameshift mutations.
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Include rescued lines (complemented with a cDNA transgene) to confirm phenotype causality.
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Monitor off-target effects via RNA-seq in CRISPR lines vs. wild-type .
What computational tools enhance epitope mapping for At3g58980 Antibody?
Integrated Workflow:
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Predict conformational epitopes using AlphaFold2-predicted structures of P0C2G1.
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Validate predictions via alanine-scanning mutagenesis coupled with ELISA .
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Cross-reference with library-on-library binding data to identify critical residues .
How to troubleshoot non-reproducible results in At3g58980 co-IP assays?
Systematic Approach:
