DRP1E Antibody
Description
Understanding DRP1E and Its Biological Roles
DRP1E is a dynamin-related protein identified in plants such as Arabidopsis thaliana and rice (Oryza sativa). Key findings include:
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Freezing Tolerance: DRP1E accumulates in sphingolipid- and sterol-enriched plasma membrane domains during cold acclimation, enhancing freezing tolerance in Arabidopsis .
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Programmed Cell Death (PCD): In rice, OsDRP1E negatively regulates PCD by controlling mitochondrial cristae morphogenesis and cytochrome c release. Mutations in OsDRP1E (e.g., E409V) impair its GTPase activity and mitochondrial localization, leading to spontaneous cell death and pathogen resistance .
Key Functional Mechanisms
Antibodies for Dynamin-Related Proteins: Focus on DRP1
While DRP1E-specific antibodies are not explicitly documented, antibodies targeting the broader DRP1 family (e.g., DRP1/Dnm1l) are widely used in research. These antibodies may cross-react with DRP1E due to conserved domains.
DRP1 Antibody Characteristics
| Antibody (Source) | Type | Applications | Reactivity | Predicted MW (kDa) | Observed MW (kDa) |
|---|---|---|---|---|---|
| Anti-DRP1 [EPR19274] (ab184247) | Recombinant Rabbit mAb | WB, ICC/IF, IHC-P, Flow Cytometry | Human, Mouse, Rat | 141, 42, 82, 86, 94 | 83, 86 |
| Anti-DRP1 [3B5] (ab56788) | Mouse mAb | IP, Flow Cyt, IHC-P | Human | 82 | 82 |
| Phospho-DRP1 (Ser616) (ab3455) | Rabbit Antibody | WB, IP, IF, Flow Cytometry | Human | 78-82 | N/A |
Notes:
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WB: Western blotting; ICC/IF: Immunocytochemistry/Immunofluorescence; IHC-P: Immunohistochemistry on paraffin sections; IP: Immunoprecipitation.
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Observed MW discrepancies may arise from post-translational modifications or isoforms.
DRP1’s Role in Mitochondrial Dynamics
DRP1 is critical for mitochondrial fission in animals, with mutations linked to neurodegenerative diseases (e.g., Alzheimer’s) and cancer . Antibodies like [EPR19274] (ab184247) are validated for detecting DRP1 in mitochondrial membranes and cytosolic fractions .
DRP1E in Plant Stress Responses
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Freezing Tolerance: DRP1E’s localization to lipid rafts in Arabidopsis plasma membranes is essential for membrane stability during cold stress .
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Pathogen Resistance: In rice, OsDRP1E mutations (e.g., E409V) disrupt mitochondrial structure, enhance cytochrome c release, and increase resistance to fungal/bacterial pathogens .
Synergistic Effects in Immunotherapy
In mammalian systems, DRP1 promotes T-cell activation and enhances PD-1/PD-L1 inhibitor efficacy in lung cancer models. For example, overexpression of DRP1 in T cells increases cytokine secretion (e.g., IFN-γ, TNF-α) and tumor cell death .
Gaps and Future Directions
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DRP1E-Specific Antibodies: No antibodies targeting DRP1E are described in the provided literature. Researchers may need to develop custom antibodies using plant-specific immunogens.
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Cross-Reactivity: DRP1 antibodies (e.g., ab56788) may bind DRP1E due to conserved domains, but validation is required.
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Therapeutic Potential: Modulating DRP1E activity could offer novel strategies for crop resilience and disease resistance in plants.
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
DRP1E Antibody targets a microtubule-associated force-producing protein. This protein is localized to the tubulo-vesicular network of the forming cell plate during cytokinesis. Furthermore, it plays a crucial role in plasma membrane maintenance and cell wall integrity, potentially influencing vesicular trafficking, polar cell expansion, and other aspects of plant growth and development. The protein also exhibits GTPase activity.
- Research on the edr3 mutant suggests a mechanistic link between salicylic acid signaling, mitochondria, and programmed cell death in plants. PMID: 16824181
Q&A
Experimental Design for DRP1E Antibody Studies
Q: How should I design experiments to study the role of DRP1E in plant immunity using DRP1E antibodies? A: To study DRP1E's role in plant immunity, consider using CRISPR-Cas9 gene editing to create overexpression and knockout lines of OsDRP1E in rice. Measure ROS generation and transcript levels of defense-related genes like PAL1, PR10, and WRKY45 in response to pathogen-associated molecular patterns (PAMPs) such as chitin. Use DRP1E antibodies for immunoblotting to confirm protein expression levels in these lines .
Antibody Selection and Validation
Q: What criteria should I use to select and validate antibodies for DRP1E research? A: When selecting antibodies for DRP1E, consider specificity, sensitivity, and cross-reactivity. Validate antibodies using techniques like Western blotting and immunofluorescence to ensure they specifically recognize DRP1E without cross-reacting with other proteins. Use controls like knockout or overexpression lines to confirm antibody specificity .
Data Interpretation and Contradiction Analysis
Q: How can I analyze and resolve contradictory data from different DRP1E antibody experiments? A: Analyze data by considering experimental conditions, antibody specificity, and biological variability. Contradictions may arise from differences in antibody affinity, sample preparation, or experimental design. Use statistical methods to assess significance and consider repeating experiments with different antibodies or techniques to validate findings .
Advanced Research Questions: Mechanistic Insights
Q: How can I use DRP1E antibodies to investigate the mechanistic role of DRP1E in mitochondrial dynamics and plant immunity? A: Use DRP1E antibodies to study protein-protein interactions by co-immunoprecipitation assays. Investigate how DRP1E interacts with other proteins involved in mitochondrial fission and plant defense pathways. This can provide insights into how DRP1E modulates mitochondrial dynamics and immune responses .
Methodological Considerations for Antibody Engineering
Q: What considerations should be taken into account when engineering antibodies for DRP1E research? A: When engineering antibodies for DRP1E, consider factors like valency, structural arrangement, and binding affinity. Optimize these parameters to achieve specific binding characteristics suitable for your research goals. Ensure manufacturability by assessing properties like expression titer, stability, and solubility .
Cross-Species Reactivity and Specificity
Q: How can I ensure that DRP1E antibodies are specific to the target species and do not cross-react with similar proteins in other species? A: To ensure specificity, use antibodies that have been validated against the target species. Perform Western blots or immunofluorescence with samples from closely related species to check for cross-reactivity. Consider using species-specific peptides for immunization to enhance specificity .
Quantitative Analysis of DRP1E Expression
Q: What methods can I use to quantify DRP1E expression levels using DRP1E antibodies? A: Use quantitative Western blotting or flow cytometry to measure DRP1E expression levels. Normalize data to housekeeping proteins or controls to ensure accurate quantification. Consider using fluorescently labeled secondary antibodies for enhanced sensitivity in flow cytometry .
Troubleshooting Common Issues
Q: What are common issues encountered when using DRP1E antibodies, and how can they be resolved? A: Common issues include non-specific binding or low signal. Resolve these by optimizing antibody dilutions, using blocking agents to reduce background, and ensuring proper sample preparation. Consider using different fixation methods or antigen retrieval techniques for immunohistochemistry .
Future Directions in DRP1E Research
Q: What are potential future directions for DRP1E antibody research in plant immunity? A: Future studies could explore the role of DRP1E in other plant-pathogen interactions or investigate how DRP1E modulates mitochondrial dynamics in response to different stressors. Additionally, developing therapeutic strategies based on DRP1E's role in plant immunity could be a promising area of research .
Example Data Table: DRP1E Antibody Validation
| Antibody Type | Species Reactivity | Application | Specificity |
|---|---|---|---|
| Monoclonal | Human, Mouse, Rat | WB, ICC, IHC | High |
| Polyclonal | Human | WB, ICC | Moderate |
| Recombinant | Human, Mouse, Rat | WB, ICC, IHC | High |
Detailed Research Findings: DRP1E and Plant Immunity
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OsDRP1E Function: OsDRP1E negatively regulates rice immunity by modulating mitochondrial dynamics. Overexpression of OsDRP1E increases susceptibility to Magnaporthe oryzae, while gene-edited plants show enhanced resistance .
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Mechanistic Insights: DRP1E's role in mitochondrial fission affects plant defense responses. Elongation of mitochondria enhances resistance to pathogens, suggesting a link between mitochondrial dynamics and immune function .
