At2g32560 Antibody
Description
Definition and Overview of At2g32560 Antibody
The At2g32560 Antibody is a polyclonal or monoclonal antibody targeting the protein encoded by the AT2G32560 gene in Arabidopsis thaliana (thale cress). This antibody is primarily used in plant biology research to study gene function, protein localization, and molecular interactions. The target protein, AT2G32560, has been implicated in circadian rhythm regulation and hypocotyl development, though its precise biochemical role remains under investigation .
Role in Plant Development
The AT2G32560 gene was identified in a study investigating circadian clock regulation and its impact on hypocotyl growth. Researchers found that decoy transgenic lines overexpressing AT2G32560 exhibited enhanced hypocotyl elongation, suggesting the gene suppresses growth under normal conditions. This phenotype was not observed in other F-box protein decoy lines, indicating AT2G32560’s unique role .
Key Findings:
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Circadian Regulation: AT2G32560 expression peaks in the morning, aligning with circadian rhythms.
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Functional Distinction: Unlike other F-box proteins (e.g., PHOTOPERIOD-INDEPENDENT EARLY FLOWERING 1), AT2G32560 does not interact with the phytochrome B (phyB) pathway, suggesting an independent regulatory mechanism .
Antibody-Based Studies
The At2g32560 Antibody enables:
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Protein Detection: Validation of gene expression via Western blot or immunoprecipitation.
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Localization Studies: Tracking AT2G32560 distribution in plant tissues (e.g., hypocotyls, leaves).
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Interaction Mapping: Identifying binding partners using co-immunoprecipitation .
Challenges:
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Limited Characterization: The antibody’s specificity and cross-reactivity in heterologous systems (e.g., non-Arabidopsis species) require validation.
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Low Signal Detection: AT2G32560 may be expressed at low levels, necessitating optimized protocols for detection .
Comparative Analysis with Related Proteins
Below is a comparison of AT2G32560 with structurally or functionally related proteins in Arabidopsis:
Note: CFH1 (CLOCK-REGULATED F-BOX WITH A LONG HYPOCOTYL 1) is the proposed name for AT2G32560 .
Future Directions
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Mechanistic Studies: Investigate how AT2G32560 regulates hypocotyl growth (e.g., via ubiquitination of growth-suppressing targets).
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Antibody Optimization: Develop recombinant antibodies with higher affinity or specificity for AT2G32560 to improve detection sensitivity .
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Evolutionary Analysis: Compare AT2G32560 orthologs in other plants to infer conserved roles in circadian regulation.
References
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Wikipedia (2025): Antibody structure and function.
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Cusabio (2025): Product details for At2g32560 Antibody (CSB-PA837296XA01DOA).
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PNAS (2024): Functional analysis of AT2G32560 in circadian regulation and hypocotyl growth.
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BioRxiv (2020): Antibody repertoire studies in plants (indirect relevance).
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NHGRI (2023): General antibody biology.
Product Specs
Constituents: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Q&A
Here’s a structured FAQ collection focused on At2g32560 Antibody for academic research, incorporating methodological guidance, experimental design considerations, and data analysis insights:
Advanced Experimental Design
How to address discrepancies in tissue-specific expression data between antibody staining and RNA-seq?
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Technical factors:
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Validation:
What strategies improve ChIP-seq signal-to-noise ratios for low-abundance At2g32560 targets?
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Optimize crosslinking conditions (e.g., 1% formaldehyde for 15 min vs. vacuum infiltration).
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Include spike-in controls (e.g., Drosophila chromatin) for normalization.
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Perform sequential ChIP (re-ChIP) to confirm co-occupancy with known interactors .
Data Interpretation & Contradictions
How to reconcile conflicting subcellular localization reports in published studies?
Why do affinity measurements vary between SPR and ITC for At2g32560 interactions?
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Surface immobilization effects in SPR may distort binding kinetics .
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Multivalent interactions: ITC measures solution-phase thermodynamics, while SPR detects surface binding.
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Validate with biolayer interferometry (BLI) under matched buffer conditions .
Methodological Optimization
How to adapt At2g32560 antibody for single-cell proteomics in root tissues?
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Use proximity ligation assays (PLA) with tyramide signal amplification.
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Validate with cell sorting + bulk LC-MS/MS from FACS-isolated cell populations .
What controls are essential for quantitative immunofluorescence in developing embryos?
| Control Type | Purpose |
|---|---|
| No-primary antibody | Autofluorescence baseline |
| Pre-immune serum | Non-specific binding |
| Competing antigen peptide (10x molar excess) | Epitope specificity |
Integration with Multi-Omics
How to correlate antibody-based protein data with transcriptomic datasets?
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Apply weighted gene co-expression network analysis (WGCNA) to identify modules linking mRNA and protein abundance.
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Account for post-transcriptional regulation using ribo-seq or tRNA profiling .
