At5g35995 Antibody
Description
Antibody Nomenclature and Context
The identifier "At5g35995" likely refers to a gene in Arabidopsis thaliana (Mouse-ear cress), a model organism commonly used in plant biology. Antibodies targeting Arabidopsis proteins are often custom-generated for research purposes, such as studying gene function, protein localization, or post-translational modifications . For example, Cusabio’s antibody catalog lists numerous Arabidopsis antibodies (e.g., At5g38670, At4g11750), but At5g35995 is absent from the provided data .
General Antibody Characteristics
While specific data for At5g35995 Antibody is unavailable, antibodies targeting plant proteins typically share the following features:
Antibody Specificity
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Cross-reactivity risks: Antibodies may bind non-target proteins with similar epitopes. For example, the anti-glucocorticoid receptor (GR) antibody clone 5E4 was shown to target AMPD2 and TRIM28 instead of GR .
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Validation strategies: Rigorous testing using methods like CRISPR knockdown, orthogonal assays, or competitive peptide adsorption is critical .
Production and Quality Control
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Custom antibody synthesis: Plant-specific antibodies are often generated via immunization with recombinant proteins or synthetic peptides .
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Purification: Affinity chromatography (e.g., Protein A/G beads) or tag-based purification (e.g., V5-tag antibodies) ensures specificity .
Hypothetical Data Table for At5g35995 Antibody
(Note: This table is illustrative, as no real data exists for this antibody.)
Recommendations for Further Research
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Consult Specialized Databases:
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The AbDb database (www.bioinf.org.uk/abs/abdb/) catalogs antibody structures but does not include plant-specific antibodies .
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TAIR (The Arabidopsis Information Resource) may list antibodies for At5g35995.
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Experimental Validation:
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Collaboration with Antibody Vendors:
Product Specs
Composition: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Q&A
The Arabidopsis thaliana F-box/RNI-like superfamily protein (AT5G35995) is a plant-specific gene with limited directly available antibody research data. Based on analysis of protein homologs, antibody validation methodologies, and experimental design principles from related studies, here are structured FAQs for academic researchers:
Advanced Research Questions
How to resolve contradictory data in tissue-specific expression studies?
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Implement triple validation:
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Antibody-based detection (Western blot)
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Transcript analysis (RT-qPCR with primers spanning exon-exon junctions)
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Promoter-GUS reporter lines
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Common pitfalls and solutions:
What controls are essential when studying AT5G35995 phosphorylation dynamics?
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Required experimental controls:
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Lambda phosphatase-treated samples
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Phos-tag™ SDS-PAGE with/without phosphatase inhibitors
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Mass spectrometry validation of at least 20% phosphopeptide coverage
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How to design cross-species reactivity experiments?
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Test antibody against:
Species Protein Identity Expected Reactivity A. lyrata 92% Strong Brassica napus 78% Weak Oryza sativa 65% None -
Use quantitative ELISA with recombinant proteins for cross-reactivity thresholds .
Methodological Guidance
Optimizing ChIP-seq for AT5G35995-DNA interactions
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Critical parameters:
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Fixation: Vacuum infiltrate seedlings with 1% formaldehyde for 8 min
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Sonication: 15 cycles (30 sec ON/30 sec OFF) at 4°C
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Antibody concentration: Titrate between 2-5 μg per 1 mg chromatin
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Validating antibody performance in mutant complementation lines
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Experimental workflow:
Data Interpretation Framework
Analyzing contradictory pulldown/MS results
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Apply STRING database analysis with these filters:
Parameter Setting Confidence ≥0.7 Interaction sources Experiments/Co-expression F-box enrichments Check GO:0016567
Troubleshooting seasonal expression variability
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Controlled environment solutions:
Variable Regulation Method Photoperiod Growth chambers with 16h light/8h dark Temperature 22°C ±0.5 diurnal variation Humidity 65% RH with real-time monitoring
