ADO1 Antibody
Description
ADO1 in Plant Systems (Arabidopsis thaliana)
Target Overview
ADO1 (Adagio Protein 1), also known as Zeitlupe (ZTL), is a blue-light photoreceptor protein in Arabidopsis thaliana involved in circadian rhythm regulation and photomorphogenesis. It functions as an F-box protein in the ubiquitin-proteasome pathway .
Antibody Characteristics
| Property | Specification |
|---|---|
| Host Species | Rabbit |
| Clonality | Polyclonal |
| Immunogen | KLH-conjugated synthetic peptide |
| Reactivity | Arabidopsis thaliana, Hordeum vulgare |
| Applications | Western blot (1:1000 dilution) |
| Molecular Weight | Predicted: 65.9 kDa |
| Supplier | Agrisera (Product AS13 2662) |
This antibody has been used to study ZTL/ADO1's role in light signaling and flowering time regulation. Its specificity for Arabidopsis makes it critical for plant chronobiology research .
Human ADO (2-Aminoethanethiol Dioxygenase)
Target Overview
Human ADO catalyzes the oxidation of cysteamine to hypotaurine and regulates thiol metabolism. It preserves oxygen homeostasis by modifying N-terminal cysteine-containing proteins .
Antibody Characteristics
| Property | Specification |
|---|---|
| Host Species | Rabbit |
| Clonality | Recombinant Monoclonal (EPR6581 clone) |
| Reactivity | Human |
| Applications | Western blot |
| Molecular Weight | ~30 kDa (observed) |
| Supplier | Abcam (Product ab134102) |
Key Functional Insights
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No reported cross-reactivity with murine homologs
Plant vs. Human ADO1 Antibodies
| Feature | Plant ADO1 Antibody | Human ADO Antibody |
|---|---|---|
| Target Function | Circadian regulation | Thiol metabolism |
| Structural Target | F-box/LOV domain protein | Non-heme iron oxygenase |
| Disease Relevance | Photoperiodism studies | Cancer, metabolic disorders |
| Commercial Availability | Agrisera | Abcam |
Plant ADO1 Studies
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ZTL/ADO1 interacts with GI (GIGANTEA) protein to stabilize TOC1 in circadian loops
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Antibody-based assays revealed diurnal expression patterns in hypocotyl growth regulation
Human ADO Research
Technical Considerations
Cross-Reactivity Challenges
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Plant ADO1 antibody shows predicted reactivity with barley (Hordeum vulgare) but not confirmed in wheat or rice
Validation Data
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
This complex targets specific clock proteins for degradation, such as APRR1/TOC1 and APRR5, but not 'GIGANTEA'. Blue light enhances the cooperative stabilization of 'GIGANTEA' and ADO1/ZTL, leading to amplification and sharpening of the expression profile of APRR1/TOC1. ADO1/ZTL interacts with ADO3, preventing the interaction of ADO3 with CDF1, further influencing the circadian clock machinery.
- Research suggests that ZTL plays a role in thermal responses and stability of the circadian clock in plants. PMID: 29172942
- Data indicate that ZEITLUPE (ZTL) E3 ubiquitin ligase-mediated protein quality control contributes to the thermal stability of the circadian clock. PMID: 29061867
- Studies reveal that ZTL forms complexes with circadian clock proteins independent of light but facilitates enhanced protein degradation in the dark. PMID: 28244872
- These data suggest the involvement of auxin and auxin-inducible genes in ZTL-mediated hypocotyl elongation. PMID: 26237185
- ZTL functions as a positive regulator in warmth-induced hypocotyl elongation under light. PMID: 26039487
- The rapid photocycle of ZTL allows detection of the day-night transition, facilitating circadian timing. The kinetic properties of ZTL reflect an evolutionary adaptation of the ZTL/FKF1/LKP2 family to function in distinct aspects of blue-light signaling. PMID: 24033190
- As the absence of either GI or ZTL compromises clock function and diminishes the protein abundance of the other, these findings highlight the critical role of their reciprocal co-stabilization for robust circadian oscillations. PMID: 24004949
- Mutations in the N terminus of ZTL interfere with binding to EBI. ZTL opposes the effects of EBI on circadian clock components during the day. PMID: 21300918
- The SCFZTL complex has been extensively characterized. PMID: 15447654
- Eleven new ztl alleles encompassing mutations in each of the ZTL protein domains have been identified. PMID: 16428597
- GIGANTEA is essential to establish and sustain oscillations of ZTL through a direct protein-protein interaction. PMID: 17704763
- TOC1/PRR3 phosphorylation-dependent interaction may protect TOC1 from ZTL-mediated degradation, resulting in an enhanced amplitude of TOC1 cycling. PMID: 18562312
Q&A
Here’s a structured FAQ collection for ADO1 antibody research, synthesized from academic literature and technical guidelines, optimized for experimental rigor and methodological clarity:
Advanced Experimental Design
How to resolve contradictory ADO1 localization data between IHC and immunofluorescence (IF)?
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Statistical approach: Apply Cohen’s kappa coefficient for inter-method agreement analysis.
What factors influence ADO1 antibody performance in flow cytometry?
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Key variables:
Methodological Challenges
How to design cross-species reactivity studies for ADO1?
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Stepwise protocol:
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Critical data: Include homology table with % identity scores.
What validation criteria apply to ADO1 antibody in ChIP-seq?
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Validation matrix:
Data Interpretation
How to distinguish true ADO1 signaling from antibody artifacts?
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Artifact identification checklist:
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Recommended tools:
ComplexHeatmap(R package) for multi-omics integration.
