At2g04740 Antibody
Description
Definition and Target Overview
The At2g04740 antibody is a specialized immunoglobulin designed to detect and bind the protein encoded by the At2g04740 gene in Arabidopsis thaliana (mouse-ear cress). This gene is annotated as a transposable_element_gene, sharing similarity with an unknown protein (TAIR:AT5G26350.1) and classified under transposable element-related functions .
Target Gene Characterization
The At2g04740 gene in Arabidopsis thaliana is associated with transposable element activity, which plays roles in genome evolution, stress responses, and epigenetic regulation. Key annotations include:
4.2. Technical Considerations
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Specificity Validation: Commercial antibodies against plant proteins frequently face cross-reactivity issues . Users should validate At2g04740 antibody specificity using knockout controls (e.g., CRISPR-edited Arabidopsis lines).
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Experimental Workflow: Pairing this antibody with techniques like chromatin immunoprecipitation (ChIP) or fluorescence in situ hybridization (FISH) could elucidate its genomic localization and interaction partners.
Future Directions
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Mechanistic Insights: Studies could explore whether At2g04740-linked proteins influence transposon silencing pathways, akin to AtSN1 or MuDR elements.
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Comparative Genomics: Cross-species analyses (e.g., with Oryza sativa transposon regulators) might reveal conserved functions.
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
Q&A
The At2g04740 antibody targets a protein encoded by the Arabidopsis thaliana gene locus AT2G04740 (Uniprot: Q9SJ85), with applications spanning functional genomics, protein interaction studies, and plant developmental biology. Below are structured FAQs addressing key research considerations, informed by antibody characterization methodologies and plant transcriptome analysis techniques .
Advanced Research Considerations
How to resolve contradictory results between Western blot and qRT-PCR data for AT2G04740 expression?
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Systematic troubleshooting protocol:
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Verify antibody specificity via 2D Western to detect charge variants (pH 3-10 gradient)
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Perform RNA-seq with 40M reads/sample to quantify splice variants potentially undetected by qRT-PCR primers
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Conduct protein turnover analysis using cycloheximide chase (100µM, 0-24hr time course)
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What controls are essential when studying AT2G04740 protein-protein interactions via co-IP?
| Control Type | Purpose | Implementation |
|---|---|---|
| Isotope-labeled competitor | Confirm binding specificity | Add 10µM 15N-labeled recombinant AT2G04740 |
| Crosslinker titration | Optimize interaction capture | Test 0.1-1.0% formaldehyde (10min fixation) |
| Mock IP | Rule out antibody leakage | Use pre-immune serum from same host species |
Methodological Challenges
How to design experiments investigating AT2G04740 phosphorylation states?
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Implement Phos-tag™ SDS-PAGE (50µM Phos-tag acrylamide) with:
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Lambda phosphatase treatment controls (30U/µl, 37°C/1hr)
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ATP depletion via 10mM 2-deoxyglucose pretreatment
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Mass spectrometry validation using TiO2 phosphopeptide enrichment
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Strategies for correlating AT2G04740 expression with heterosis traits in hybrids
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Combine antibody-based quantification with transcriptome profiling:
