CTG10 Antibody
Description
Molecular Characterization of CTG10 Antibody
The CTG10 antibody was generated through peptide immunization and affinity purification for precise detection in plant studies .
Key specifications:
| Parameter | Details |
|---|---|
| Antigen peptide | MAYLSFKSNMERTPRESNTPC (21-mer with C-terminal cysteine) |
| Host species | New Zealand White rabbit |
| Purification method | Affinity chromatography using agarose-linked peptide |
| Applications | Western blotting, coimmunoprecipitation (Co-IP), protein quantification |
Functional Insights from CTG10-PIF1 Interaction Studies
CTG10 regulates PHYTOCHROME-INTERACTING FACTOR 1 (PIF1) stability through direct interaction, as demonstrated by:
Yeast Two-Hybrid Binding Assay
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Full-length PIF1 showed strongest interaction with CTG10 (β-galactosidase activity = 12.4 ± 1.2 units)
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Binding efficiency of PIF1 fragments:
| PIF1 Domain | Relative Interaction Strength (%) |
|---|---|
| N-terminal (1-180 aa) | 34.7 ± 3.1 |
| C-terminal (181-478 aa) | 22.9 ± 2.8 |
| bHLH region (100-160 aa) | 68.5 ± 4.2 |
Protein Stability Regulation
Western blot analyses using CTG10 antibody revealed:
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CTG10 overexpression (CTG10-OE) reduced PIF1 levels by 62% under red light vs wild type
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Proteasome inhibitor (bortezomib) treatment increased PIF1 accumulation by 3.8-fold in dark-grown seedlings
Technical Validation Metrics
The antibody's performance was rigorously validated:
Western blot sensitivity:
| Sample Type | Detection Limit |
|---|---|
| Crude plant extract | 5 µg total protein |
| Affinity-purified CTG10 | 0.1 ng |
Co-IP efficiency:
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Immunoprecipitated CTG10-GFP complexes contained PIF1 at 18.7 ± 2.3 ng/mg total protein
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Light treatment reduced CTG10-PIF1 interaction by 74% compared to dark conditions
Research Applications
The CTG10 antibody has enabled critical discoveries in plant photomorphogenesis:
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Mechanistic studies
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Quantified PIF1 degradation kinetics under different light spectra
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Mapped interaction domains through truncation mutants
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Developmental regulation
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Technical applications
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Enabled simultaneous detection of CTG10 and tubulin (loading control) in multiplex Western blots
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Facilitated longitudinal protein tracking through quantitative densitometry
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Product Specs
Constituents: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Target Background
KEGG: ath:AT4G19330
STRING: 3702.AT4G19330.1
Q&A
Here’s a structured collection of FAQs for researchers working with CTG10 antibodies, organized by research complexity and supported by experimental data:
What experimental designs address conflicting CTG10 quantification results across assays?
Resolution strategy:
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Normalize data to tubulin or total protein (Ponceau S staining) to account for loading variability .
How to optimize CTG10 antibody performance in co-immunoprecipitation (co-IP)?
Protocol refinement:
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Pre-clear lysates with Protein A/G beads to reduce nonspecific binding.
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Use 200 µM bortezomib during extraction to stabilize ubiquitinated complexes .
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Validate interactions via reciprocal IP (e.g., anti-GFP pulldown in CTG10-GFP transgenic lines) .
What advanced techniques confirm CTG10’s role in seed germination signaling?
Integrated workflow:
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Yeast two-hybrid screening: Identify CTG10-PIF1 interactions using full-length PIF1 constructs (stronger binding than truncated versions) .
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Functional assays: Compare germination rates in CTG10-overexpressing vs. knockout Arabidopsis lines under red light (100 µmol/m²) .
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Cytokine profiling: Measure IL-2, IL-10, and Th2 cytokines (e.g., IL-4) in vivo to link CTG10 to immunosuppressive pathways .
How to engineer single-chain variable fragments (scFvs) targeting CTG10?
Biophysical optimization:
How to assess CTG10 antibody stability in longitudinal in vivo studies?
Pharmacokinetic analysis:
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Administer 10 mg/kg doses (vs. 3 mg/kg) to maintain plasma N+W-CTGF levels >14 days post-infusion .
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Monitor clearance rates using ELISA:
