SRO1 Antibody
Description
SRO1 Antibody Functional Context
SRO1 is a nuclear protein that interacts with transcription factors involved in oxidative stress responses and hormonal signaling pathways. Key characteristics include:
Research Applications
SRO1 antibodies have been critical in elucidating:
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Stress Adaptation: SRO1 regulates transcriptional networks during drought, salinity, and UV exposure by interacting with AP2/ERF, NAC, and bHLH transcription factors .
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Developmental Regulation: SRO1 modulates root architecture and leaf senescence via auxin and jasmonate signaling pathways .
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DNA Repair Mechanisms: SRO1 collaborates with RCD1 to mitigate reactive oxygen species (ROS)-induced DNA damage .
Interaction Partners
A yeast two-hybrid screen using SRO1 antibodies identified:
| Transcription Factor Family | Number of Interacting Partners | Biological Process |
|---|---|---|
| AP2/ERF | 14 | Abiotic stress responses |
| NAC | 9 | Senescence and cell death |
| bHLH | 6 | Light signaling and stomatal development |
Phenotypic Analysis
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Single Mutants: sro1 mutants show no visible developmental defects under standard conditions .
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Double Mutants: rcd1 sro1 mutants exhibit stunted growth, chlorosis, and reduced fertility, indicating synergistic roles in stress tolerance .
Technical Validation
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Western Blot: SRO1 antibodies detect a ~55 kDa band in wild-type Arabidopsis extracts, absent in sro1 knockout lines .
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Subcellular Localization: Immunofluorescence confirms SRO1’s nuclear localization, consistent with its role in transcriptional regulation .
Limitations and Future Directions
Current research gaps include:
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Structural details of SRO1-antigen binding interfaces.
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Conservation of SRO1 homologs in crop species.
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Therapeutic potential in agricultural biotechnology (e.g., engineered stress-resistant plants).
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
- Both RCD1 and SRO1 are essential for proper cell differentiation. PMID: 21172813
- RCD1 and SRO1 are required for the appropriate development of Arabidopsis, exhibiting a degree of functional redundancy. PMID: 19548978
- Research indicates that RCD1 and SRO1 exhibit at least partial redundancy and are essential genes for plant development. PMID: 19625634
Q&A
Basic Research Questions
What experimental approaches confirm Sro1's role in oxidative stress response?
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Methodological framework:
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Use Δsro1 knockout strains alongside wild-type Schizosaccharomyces pombe
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Treat with stressors (e.g., 40 μL CSE, 5 mM hydroxyurea, 42°C heat shock)
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Quantify ROS levels via H2-DCFDA staining and flow cytometry at 0/30/60/120 min intervals
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Compare cell-cycle progression using DNA content analysis (1N/2N/4N peaks)
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Key data table:
| Stress Condition | Wild-Type ROS Profile | Δsro1 ROS Profile | Checkpoint Activation Delay |
|---|---|---|---|
| CSE Exposure | Transient peak → decline | Sustained elevation | S-phase delay (2h) |
| Heat Shock | Gradual accumulation | 40% reduced levels | G2/M prolongation |
| Hydroxyurea | Biphasic response | Inverted trajectory | Premature S-phase exit |
How to validate Sro1 antibody specificity in fission yeast models?
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Validation pipeline:
Advanced Research Challenges
How to reconcile contradictory ROS modulation data across stress conditions?
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Resolution strategy:
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Establish standardized treatment protocols:
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CSE: 40 μL dose for 60 min
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Heat shock: 42°C with pre-acclimation phase
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Implement single-cell ROS imaging to reduce population averaging artifacts
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Profile mitochondrial membrane potential (ΔΨm) using TMRE staining
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What mechanistic models explain Sro1's dual role in ROS elevation/scavenging?
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Experimental design:
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Hypothesis testing framework:
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Model A: Subcellular localization shifts under different stresses
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Model B: Post-translational modifications alter enzyme binding
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Key experiments:
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How to optimize cell-cycle synchronization for Sro1 interaction studies?
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Protocol optimization:
Synchronization Method G1 Yield S-Phase Purity Compatibility with Sro1 Studies Lactose gradient 82% ±15% High (no chemical interference) Hydroxyurea block 68% ±8% Moderate (alters ROS baseline) Elutriation 91% ±5% Optimal for checkpoint analysis
What computational tools integrate Sro1's stress-specific regulatory networks?
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Systems biology approach:
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Build Boolean network incorporating:
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SAPK pathway inputs (sty1/atf1)
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Cell-cycle regulators (rad3/chk1/cds1)
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Metabolic outputs (ethanol utilization efficiency)
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Validate predictions using:
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Methodological Considerations
How to address pathway cross-talk in Sro1 antibody-based assays?
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Interference mitigation:
What controls are essential for Sro1 colocalization studies?
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Validation matrix:
Control Type Purpose Acceptable Threshold Isotype-matched IgG Background binding <5% cellular signal Δsro1 strain Antibody specificity 0% detectable signal Fixed vs live-cell imaging Artifact discrimination ≤15% signal variance Stress-naïve cells Baseline localization pattern Nuclear:cytosol 3:1
