At1g61730 Antibody
Description
Overview of At1g61730 Protein
The At1g61730 protein is a DNA-binding storekeeper protein-related transcriptional regulator identified in Arabidopsis thaliana. Key characteristics include:
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Function: Regulates transcription by interacting with chromatin or DNA-binding components.
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Expression: Detected in both wild-type (WT) and mutant (al6) plant genotypes .
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Peptide Identification: Contains 6 unique peptides in proteomic analyses, confirming its presence in chromatin-enriched fractions .
| Locus | Name | Function | Unique Peptides | Genotype |
|---|---|---|---|---|
| At1g61730.1 | DNA-binding storekeeper protein-related transcriptional regulator | Regulation of transcription | 6 | WT; al6 |
Antibody Function and Mechanism
Antibodies targeting At1g61730 are designed to:
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Detect Protein Localization: Track At1g61730 in nuclear or chromatin-associated complexes.
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Study Protein Interactions: Identify binding partners via co-immunoprecipitation (e.g., DEK3 or SCC3 in chromatin remodeling studies ).
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Analyze Gene Expression: Investigate transcriptional regulation in response to environmental or genetic cues.
Antibody Design Considerations
While specific data on At1g61730 antibodies are unavailable, general principles apply:
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Epitope Selection: Target conserved regions (e.g., DNA-binding domains) to ensure specificity.
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Validation: Use Western blotting or immunoprecipitation to confirm binding .
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Cross-Reactivity: Minimize binding to homologous proteins (e.g., other storekeeper proteins).
Chromatin and Transcriptional Regulation
At1g61730 is implicated in chromatin architecture studies. For example:
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ChEP-P Method: Chromatin enrichment proteomics identified At1g61730 alongside proteins like ALBA1 (chromatin structure) and GRF1 (transcription regulation) .
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DEK3 Interactions: Proteins regulating DNA topology (e.g., DEK3) often associate with transcriptional regulators, suggesting At1g61730 may modulate chromatin accessibility .
Potential Applications
Challenges and Future Directions
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Limited Data: No peer-reviewed studies explicitly describe At1g61730 antibodies.
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Technical Hurdles:
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Antigen Complexity: At1g61730’s role in chromatin may require antibodies with high affinity to nuclear proteins.
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Cross-Reactivity: Potential overlap with homologous storekeeper proteins.
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Opportunities:
Product Specs
Composition: 50% Glycerol, 0.01M PBS, pH 7.4
Q&A
Here’s a curated collection of FAQs for researchers working with the At1g61730 Antibody in academic settings, synthesized from peer-reviewed methodologies and experimental design principles:
What experimental applications are best suited for At1g61730 Antibody?
Key applications:
How can I resolve non-specific bands in Western blot experiments?
Advanced troubleshooting:
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Buffer optimization: Test alternative blocking agents (e.g., 5% BSA vs. non-fat milk) to reduce background .
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Cross-reactivity checks: Use KO lysates to identify non-target proteins and validate with independent antibodies targeting the same protein .
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Antibody dilution gradients: Titrate from 1:500 to 1:2000 to balance signal-to-noise ratios.
What strategies mitigate batch-to-batch variability in long-term studies?
Best practices:
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Lot validation: Compare new antibody lots against previous batches using standardized lysates and protocols .
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Reference standards: Preserve aliquots of positive-control lysates for longitudinal comparisons.
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Multiplex validation: Pair At1g61730 Antibody with housekeeping protein antibodies (e.g., actin) to normalize signal drift .
How do I integrate At1g61730 Antibody into high-dimensional flow cytometry panels?
Spectral cytometry workflow:
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Panel design: Assign At1g61730 to a high-sensitivity channel (e.g., BV421) to avoid spectral overlap with CD3/CD45 markers .
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Data analysis: Use Leiden clustering in Python to identify rare populations (e.g., <0.1% abundance) and UMAP for visualization .
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Cross-validation: Compare flow results with single-cell RNA-seq data to confirm protein-RNA expression correlations.
How should I address contradictions between published studies using this antibody?
Critical analysis framework:
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Technical audit: Compare methods for lysate preparation, fixation, and antibody dilution across conflicting studies .
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Orthogonal validation: Confirm target engagement via CRISPR interference or siRNA knockdown .
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Meta-analysis: Aggregate data from public repositories (UniProt: Q9SYA9) to assess consensus epitope recognition patterns .
