daf-25 Antibody
Description
Molecular Characterization of daf-25
daf-25 encodes the Caenorhabditis elegans ortholog of mammalian Ankmy2, a protein containing three ankyrin repeats and a zinc finger MYND domain . This protein localizes to sensory cilia and regulates the trafficking of membrane-bound guanylyl cyclases (GCs), which are critical for cyclic GMP (cGMP) signaling .
Key Features of DAF-25:
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Domain Architecture:
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Ankyrin repeats (protein-protein interactions)
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MYND domain (zinc finger involved in transcriptional regulation)
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Cellular Localization: Sensory cilia in chemosensory neurons .
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Functional Role: Facilitates ciliary localization of GCs, such as DAF-11, enabling cGMP-dependent signaling for chemosensation and developmental decisions .
Functional Insights from Genetic Studies
DAF-25 is essential for proper ciliary function and interacts with components of the insulin/IGF-1 and TGF-β signaling pathways.
Genetic Interactions and Phenotypic Rescue:
Key Findings:
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Dauer Formation: daf-25 mutants exhibit constitutive dauer larva formation under non-stress conditions, implicating its role in environmental sensing .
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cGMP Pathway: Exogenous cGMP rescues daf-25 mutant phenotypes, confirming its upstream role in cGMP signaling .
Implications for Human Disease
DAF-25’s mammalian counterpart, Ankmy2, binds retinal guanylyl cyclase GC1, suggesting a conserved role in vision . Dysregulation of Ankmy2 may contribute to:
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Retinal Degeneration: Altered GC1 trafficking could impair phototransduction.
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Ciliopathies: Defects in ciliary GC localization linked to obesity and Bardet-Biedl syndrome .
Research Applications of daf-25 Antibody
While commercial antibodies specific to C. elegans DAF-25 are not explicitly detailed in available sources, polyclonal or monoclonal antibodies generated for research purposes enable:
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Localization Studies: Immunofluorescence to track DAF-25 in sensory cilia .
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Western Blotting: Detection of DAF-25 in protein lysates from C. elegans mutants .
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Functional Assays: Blocking DAF-25 to study ciliary GC trafficking defects .
Future Directions
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Therapeutic Targeting: Modulating Ankmy2 in retinal or metabolic disorders.
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Mechanistic Studies: Elucidating how MYND domains regulate GC trafficking.
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
Q&A
The following FAQs address key scientific considerations for researchers working with Dual Action Fab (DAF) antibodies, focusing on academic research rigor and methodological approaches. Content is synthesized from peer-reviewed studies, patent filings, and biotechnology protocols.
What experimental workflows validate dual-specific binding in vitro?
Methodological Protocol
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Surface plasmon resonance (SPR):
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Flow cytometry co-staining:
Data Conflict Resolution
If SPR and cell assays disagree:
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Check for epitope masking in cell membranes vs. recombinant proteins.
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Perform competitive ELISA with soluble antigens (IC50 ratio <2.0 suggests true dual binding) .
How to optimize DAF antibodies for in vivo therapeutic efficacy?
Pharmacokinetic Considerations
| Parameter | Target Range | Rationale |
|---|---|---|
| Serum half-life | ≥7 days | Match IgG’s Fc-mediated recycling |
| Tumor penetration | ≥5% injected dose/g | Optimize isoelectric point (pI 6.5–7.5) via histidine substitutions |
Combination Therapy Design
What computational models predict cross-reactivity risks for novel DAF designs?
Advanced Modeling Workflow
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Epitope binning: Cluster antigens using AlphaFold-Multimer-predicted interfaces.
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Molecular dynamics (MD): Simulate 100-ns trajectories to quantify paratope flexibility (RMSF >2.5 Å indicates instability) .
Validation Pipeline
| Step | Success Criterion |
|---|---|
| Phage display panning | Enrichment ratio ≥1e5 vs. non-target antigen |
| Deep mutational scanning | <5% variants show >10-fold affinity loss |
How to troubleshoot non-specific signaling activation in ErbB-targeting DAFs?
Mechanistic Analysis
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Phosphoproteomics: Compare ErbB2 phosphorylation in BT-474 cells (high ErbB2) vs. MCF-7 (low ErbB2) after DAF treatment:
Contradictory Data Protocol
If unexpected signaling occurs:
