FDM3 Antibody
Description
FDM3: Molecular and Functional Overview
FDM3 (Factor of DNA Methylation 3) is one of five SGS3-like proteins in Arabidopsis (At3G12550) that play roles in RdDM, a plant-specific epigenetic silencing pathway .
Key Features
| Property | Description |
|---|---|
| Protein Family | SGS3-like proteins (subfamily 2) |
| Domains | Four conserved motifs: XH, XS, coiled-coil, and zinc finger (VWA domain) |
| Molecular Weight | ~110 kDa (predicted) |
| Expression | Co-expressed with RdDM machinery components (e.g., Pol V, AGO4) |
Genetic Redundancy
FDM3 acts redundantly with other SGS3-like proteins:
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FDM1/FDM2: Subfamily 3 proteins with overlapping roles in RdDM .
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IDN2: Subfamily 2 protein required for scaffold RNA production .
DNA Methylation Defects in Mutants
Double mutants involving fdm3 show stronger DNA hypomethylation than single mutants :
| Locus | WT Methylation (%) | fdm3-1 (%) | fdm1-1 fdm3-1 (%) |
|---|---|---|---|
| AtSN1 | 85 | 82 | 45 |
| IGN5 | 78 | 75 | 32 |
| 5S rDNA | 90 | 88 | 60 |
Functional Overlap
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Genetic Interactions:
FDM3 Antibody: Current Status
No commercially available antibodies specifically targeting FDM3 are documented in the provided literature. Research on FDM3 relies on genetic tools (e.g., T-DNA insertion lines, transcript analysis) .
Comparison with DF3 Antibody
The DF3 monoclonal antibody (unrelated to FDM3) targets a 300 kDa mammary epithelial antigen and has clinical applications in breast cancer diagnostics .
Future Directions
While FDM3 itself is critical in plant epigenetics, the absence of FDM3-specific antibodies limits mechanistic studies. Development of such reagents could enable:
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Subcellular localization studies
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Protein-protein interaction assays
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Tissue-specific expression profiling
Product Specs
Composition: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Target Background
Q&A
Given the lack of specific information on "FDM3 Antibody" in the search results, I will provide a general set of FAQs for researchers on antibodies, focusing on experimental design, data analysis, and methodological approaches. These FAQs are designed to reflect the depth of scientific research and distinguish between basic and advanced research questions.
Answer:
Validating an antibody involves several steps:
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Positive and Negative Controls: Use known positive and negative controls to assess antibody specificity and sensitivity. This can include cell lines or tissue samples with variable expression levels of the target protein .
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Western Blot and Immunofluorescence: Perform Western blotting to check for specific banding patterns and immunofluorescence to verify cellular localization .
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Orthogonal Controls: Use knockout cell lines or RNA interference to confirm specificity .
Answer:
Conflicting data can arise from differences in assay conditions or antibody specificity. To resolve this:
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Standardize Assay Conditions: Ensure that all assays are performed under the same conditions (e.g., buffer, temperature).
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Use Multiple Antibodies: Validate findings with multiple antibodies targeting different epitopes of the same protein.
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Consider Post-Translational Modifications: Differences in protein modifications might affect antibody binding .
Answer:
To ensure specificity and minimize cross-reactivity:
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Epitope Mapping: Identify the specific epitope recognized by the antibody to understand potential cross-reactivity .
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Use of Knockout Controls: Validate specificity using knockout cell lines or tissues lacking the target protein .
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Competitive Binding Assays: Perform competitive binding assays to assess specificity against closely related proteins .
Answer:
To design antibodies with specific profiles:
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Phage Display Technology: Utilize phage display experiments to select antibodies against specific ligands .
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Computational Modeling: Employ computational models to predict and design antibodies with desired binding modes .
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High-Throughput Sequencing: Use sequencing data to analyze antibody libraries and identify clones with desired specificity .
Answer:
Common challenges include:
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Antibody Specificity: Addressed through rigorous validation and use of orthogonal controls .
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Scalability and Reproducibility: Ensure consistent conditions across experiments and consider using recombinant antibodies for better reproducibility .
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Data Sharing and Transparency: Encourage open sharing of antibody performance data to improve community-wide validation efforts .
Answer:
Future directions include:
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Recombinant Antibody Technology: Offers improved specificity and reproducibility compared to traditional monoclonal antibodies .
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Computational Antibody Design: Enhances the ability to predict and customize antibody specificity .
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Collaborative Efforts: Encourage partnerships between researchers and vendors to improve antibody validation and availability .
Data Table Example: Antibody Validation Methods
| Method | Description | Advantages |
|---|---|---|
| Western Blot | Assesses protein expression and specificity | Provides molecular weight information |
| Immunofluorescence | Visualizes protein localization | Offers spatial resolution of protein distribution |
| Knockout Controls | Validates specificity by absence of signal in knockout samples | Ensures target specificity |
