At5g46170 Antibody
Description
Key Features:
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Immunogen: Recombinant Arabidopsis thaliana At5g46170 protein.
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Host: Rabbit.
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Reactivity: Specific to Arabidopsis thaliana.
Recombinant Protein Forms and Applications
The At5g46170 protein is available in multiple recombinant forms, enabling diverse experimental approaches:
| Source Organism | Conjugation | Applications |
|---|---|---|
| Saccharomyces cerevisiae (yeast) | None | Protein expression, purification |
| E. coli | AviTag-Biotinylated | Pull-down assays, protein labeling |
| Baculovirus-infected cells | None | Structural studies, protein folding |
| Mammalian cells | None | Native protein conformation analysis |
These forms allow researchers to study the protein’s function in ubiquitin-mediated degradation pathways or its interactions with other components of the SCF (Skp1-Cullin-F-box) complex .
Role of At5g46170 in Plant Pathways
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F-box Proteins: At5g46170 belongs to the F-box protein family, which recruits substrates to the SCF complex for ubiquitination and proteasomal degradation .
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Potential Applications:
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Protein Degradation: The antibody can track substrate turnover regulated by At5g46170.
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Stress Responses: F-box proteins often mediate stress responses (e.g., drought, pathogens); the antibody may aid in studying these pathways.
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Experimental Validation
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Western Blot: Used to confirm the presence of At5g46170 in Arabidopsis extracts.
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ELISA: Quantifies protein levels in developmental or stress-induced samples .
Comparative Analysis with Other Antibodies
The At5g46170 Antibody shares structural and functional parallels with other polyclonal antibodies but is tailored for plant-specific studies:
| Feature | At5g46170 Antibody | General Polyclonal Antibodies |
|---|---|---|
| Specificity | Arabidopsis thaliana | Broad species reactivity |
| Epitope Diversity | Multiple epitopes | Variable |
| Primary Use | Plant protein studies | General antigen detection |
Future Directions
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Therapeutic Targets: While not directly therapeutic, insights into At5g46170’s role in protein degradation could inform strategies for manipulating plant stress responses.
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Cross-Species Studies: Potential applications in studying F-box protein homologs in other plants or model organisms.
Product Specs
**Constituents:** 50% Glycerol, 0.01M PBS, pH 7.4
Q&A
Here’s a structured FAQ collection for researchers working with the At5g46170 Antibody, focusing on academic research applications and methodological rigor:
How to resolve conflicting results in protein abundance measurements using At5g46170 Antibody?
Advanced Research Focus
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Technical variability: Normalize signals using housekeeping proteins (e.g., actin) across replicates.
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Epitope masking: Pre-treat samples with protein unfolding buffers (e.g., 6M urea) to expose cryptic epitopes .
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Data integration: Combine antibody-derived protein quantification with transcriptomic data (e.g., RNA-seq) to identify post-transcriptional regulation patterns .
What are optimal conditions for co-immunoprecipitation (Co-IP) assays using At5g46170 Antibody?
Methodological Guidance
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Lysis buffer: Use RIPA buffer supplemented with 1% Triton X-100 and protease inhibitors.
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Crosslinker choice: For transient interactions, apply formaldehyde crosslinking (1% for 10 min) .
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Elution strategy: Compete with 0.1M glycine (pH 2.5) to preserve protein complexes for downstream MS analysis.
How to design a CRISPR knockout line for validating At5g46170 Antibody specificity?
Experimental Design
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Target exons: Design sgRNAs against exon 2–3 to disrupt the F-box domain (amino acids 50–150).
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Phenotypic validation: Screen T2 lines for developmental defects (e.g., delayed flowering) linked to F-box protein dysfunction .
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Antibody testing: Confirm loss of signal in Western blots and immunofluorescence assays .
What computational tools can predict interaction partners of the At5g46170 F-box protein?
Advanced Data Analysis
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Network modeling: Use STRING (https://string-db.org) with Arabidopsis interactome data.
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Domain analysis: Screen for proteins with Leu-rich repeats (LRR) or SKP1-like domains using InterProScan.
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Validation workflow:
How to optimize multiplex imaging with At5g46170 Antibody in tissue sections?
Technical Optimization
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Antibody compatibility: Use Alexa Fluor® 488 (green) for At5g46170 and far-red secondary antibodies (e.g., Cy5) for co-staining.
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Autofluorescence reduction: Treat tissues with 0.1% Sudan Black B in 70% ethanol for 30 min .
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Signal quantification: Apply deconvolution algorithms (e.g., Huygens Software) to resolve subcellular structures.
What statistical approaches address batch effects in large-scale At5g46170 protein studies?
Data Contradiction Mitigation
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ComBat correction: Harmonize protein abundance data across experimental batches using the sva R package .
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Replicate design: Include biological replicates from independently grown plant cohorts.
| Batch Effect Source | Correction Strategy |
|---|---|
| Day-to-day variation | ComBat harmonization |
| Tissue harvesting time | Z-score normalization |
