ATXR2 Antibody
Description
ATXR2 Protein Overview
ATXR2 (ARABIDOPSIS TRITHORAX-RELATED 2) is a histone lysine methyltransferase belonging to the Trithorax Group (TrxG) in plants. It catalyzes the deposition of H3K36me3 marks, facilitating transcriptional activation during cell fate transitions like callus formation and shoot/root organogenesis . Key functions include:
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Regulating auxin-cytokinin interactions during shoot regeneration by interacting with ARR1 to activate type-A ARR genes .
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Promoting dedifferentiation by depositing H3K36me3 at LBD gene promoters, enabling callus formation .
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Controlling root identity through interactions with ARF7/19 and LBD16/29 during lateral and adventitious root formation .
Plant Studies (ATXR2)
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Callus Formation: ATXR2-deficient Arabidopsis mutants show reduced callus formation, while overexpression enhances it .
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Gene Targets: Binds promoters of LBD16/29 and ARR5/7, modulating cytokinin signaling and WUSCHEL activation .
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Developmental Role: Essential for lateral root initiation and adventitious root formation .
Human Studies (Ataxin-2)
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Disease Association: PolyQ expansions in ATXN2 correlate with ALS and neurodegeneration .
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Molecular Mechanisms: Facilitates stress granule assembly and interacts with TDP-43, a protein implicated in ALS pathology .
Comparative Analysis of ATXR2 and Ataxin-2
Future Directions
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- ATXR2, a histone lysine methyltransferase, promotes the accumulation of histone H3 proteins trimethylated at lysine 36 (H3K36me3) during callus formation. This accumulation contributes to the early stages of cellular dedifferentiation by activating LATERAL ORGAN BOUNDARIES DOMAIN (LBD) genes. PMID: 29184030
Q&A
Experimental Design for Studying ATXR2 Function
Q: How can I design an experiment to study the role of ATXR2 in plant regeneration using ATXR2 antibodies? A: To study ATXR2's role in plant regeneration, you can use ATXR2 antibodies in immunoprecipitation assays to identify interacting proteins involved in shoot organogenesis. For instance, co-immunoprecipitation with ARR1 can help elucidate the auxin-cytokinin interaction during shoot regeneration . Additionally, use Western blotting to assess ATXR2 expression levels in callus formation and root organogenesis .
Data Analysis and Contradiction Resolution
Q: How do I resolve contradictions in data when using ATXR2 antibodies in different experimental conditions? A: When encountering data contradictions, ensure that the ATXR2 antibody specificity is validated across different experimental conditions. Use controls such as isotype controls to rule out non-specific binding . Also, consider the impact of environmental factors and genetic backgrounds on ATXR2 expression and activity .
Advanced Research Questions: Epigenetic Regulation
Q: What role does ATXR2 play in epigenetic regulation during plant cell dedifferentiation, and how can this be studied using ATXR2 antibodies? A: ATXR2 deposits H3K36me3 marks at LBD gene promoters, facilitating cellular dedifferentiation . To study this, use chromatin immunoprecipitation (ChIP) assays with ATXR2 antibodies to analyze histone modifications at specific gene loci. This can provide insights into how ATXR2 regulates gene expression during callus formation .
Methodological Considerations for Antibody Selection
Q: What factors should I consider when selecting an ATXR2 antibody for research purposes? A: When choosing an ATXR2 antibody, consider the specificity, sensitivity, and cross-reactivity. Ensure the antibody is validated for your specific application (e.g., Western blot, immunoprecipitation) and species. Also, check if the antibody has been tested in similar experimental conditions to yours .
Optimization of Antibody Concentration
Q: How do I optimize the concentration of ATXR2 antibodies for different applications? A: Start with the recommended concentration provided by the manufacturer and optimize it based on your experimental conditions. Factors such as protein expression levels, sample preparation, and detection system can affect the optimal concentration. Perform titration experiments to find the best concentration for your specific application .
Interpretation of Results in Different Plant Tissues
Q: How do I interpret the results of ATXR2 antibody staining in different plant tissues? A: When interpreting results from different plant tissues, consider the developmental stage and tissue type. ATXR2 plays roles in both shoot and root organogenesis, so its expression might vary significantly across tissues . Use controls and comparative analyses to understand the functional implications of ATXR2 expression in each tissue.
Combining ATXR2 Antibodies with Other Techniques
Q: How can I combine ATXR2 antibodies with other molecular biology techniques to enhance research outcomes? A: Combine ATXR2 antibodies with techniques like qRT-PCR to analyze gene expression changes, or with ChIP-seq to map genome-wide histone modifications. This multi-approach strategy can provide comprehensive insights into ATXR2's role in plant development and regeneration .
Troubleshooting Common Issues
Q: What are common issues encountered when using ATXR2 antibodies, and how can they be resolved? A: Common issues include non-specific binding and low signal intensity. Resolve these by optimizing antibody concentrations, using appropriate blocking solutions, and ensuring proper sample preparation. Also, validate the specificity of the antibody by using isotype controls .
Example Data Table: ATXR2 Antibody Optimization
| Antibody Concentration (µg/mL) | Signal Intensity (Western Blot) | Background Noise |
|---|---|---|
| 0.1 | Low | High |
| 0.5 | Medium | Medium |
| 1.0 | High | Low |
In this example, an antibody concentration of 1.0 µg/mL provides the best signal-to-noise ratio for Western blot analysis.
