At5g02060 Antibody
Description
Potential Applications
While no peer-reviewed studies explicitly cite the At5g02060 Antibody, its design suggests utility in:
| Application | Description |
|---|---|
| Immunoblotting (WB) | Detection of Q9LZM5 protein in lysates |
| Immunoprecipitation (IP) | Purification of protein complexes |
| Immunohistochemistry | Localization of Q9LZM5 in plant tissues |
| Gene Expression Studies | Quantifying protein levels during growth |
Antibody Specificity
The antibody’s specificity is inferred from its targeting of the At5g02060 gene product. Cross-reactivity with homologous proteins in other species is not explicitly reported but may require validation.
Research Gaps and Future Directions
To date, no published studies directly validate the At5g02060 Antibody’s performance in experimental setups. Future research could focus on:
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Epitope Mapping: Identifying the antibody’s binding site on Q9LZM5.
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Functional Studies: Linking Q9LZM5 to metabolic pathways or stress responses in Arabidopsis.
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Comparative Analysis: Validating cross-reactivity with orthologs in model plants (e.g., rice, maize).
Availability and Procurement
The antibody is commercially available through Cusabio (Table 1). Researchers should confirm lot-specific performance and dilution protocols prior to use.
Product Specs
Constituents: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Target Background
KEGG: ath:AT5G02060
UniGene: At.50418
Q&A
Here’s a structured collection of FAQs tailored to researchers working with the Arabidopsis thaliana AT5G02060 antibody, synthesized from cross-disciplinary methodologies in antibody research and plant molecular biology:
What experimental designs address conflicting localization data for AT5G02060 across studies?
Advanced approach:
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Conduct compartment-specific fractionation (nuclear/cytoplasmic/membrane) followed by immunoblotting.
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Use confocal microscopy with organelle markers (e.g., chloroplast: CAB3-RFP; nucleus: H2B-YFP) in stably transformed Arabidopsis.
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Analyze tissue-specific expression via promoter-GUS fusions combined with antibody staining .
Contradiction resolution framework:
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Compare antibody clones (polyclonal vs. monoclonal)
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Verify fixation/permeabilization protocols (e.g., Triton X-100 concentration)
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Assess developmental stage-dependent expression
How to optimize AT5G02060 antibody concentration for multiplexed imaging?
Titration strategy:
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Test 4-fold dilutions (0.16–10 µg/mL) in PBS-T with 1% BSA
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Use computational tools (e.g., CODEX/IBEX platforms) to quantify signal-to-background ratios
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Reference: Oligo-conjugated antibody studies show saturation typically occurs at 0.62–2.5 µg/mL
Multiplex panel optimization:
| Parameter | Recommended Range | Rationale |
|---|---|---|
| Incubation time | 2–4 hr RT | Minimizes non-specific binding |
| Blocking agent | 5% skim milk + 0.1% Tween-20 | Reduces plant phenolic interference |
| Imaging order | AT5G02060 first | Prevents epitope damage from harsh elution |
What structural features of AT5G02060 impact antibody-antigen binding?
Epitope analysis:
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Predict disordered regions using IUPred2A; linear epitopes often reside in these regions
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Compare homology models (AlphaFold2) with cross-reactive species (e.g., Brassica napus orthologs)
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For conformational epitopes: Perform limited proteolysis-mass spec to map protected regions
Key biophysical parameters:
| Feature | Impact on Antibody Performance |
|---|---|
| Post-translational modifications (e.g., phosphorylation at YXX) | May require non-denaturing WB conditions |
| Quaternary structure (monomer vs. dimer) | Affects immunoprecipitation efficiency |
| Redox-sensitive cysteines | Include 2 mM TCEP in lysis buffer |
How to interpret AT5G02060 antibody signals in mutant complementation lines?
Quantitative framework:
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Normalize signals to housekeeping proteins (e.g., ACTIN8) across biological replicates (n ≥ 5)
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Use RNAi knockdowns to establish linear dynamic range (10–100 µg total protein loaded)
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For partial complementation: Calculate % wild-type signal using densitometry (ImageJ FIJI)
Advanced controls:
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Spiked-in recombinant AT5G02060 (5–500 ng) to validate detection limits
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Isothermal titration calorimetry to measure binding affinity (K<sub>d</sub>)
What computational tools predict AT5G02060-antibody interactions for rational design?
AI-driven solutions:
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Use MAGE-like frameworks to generate synthetic heavy/light chain pairs against predicted epitopes
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Molecular dynamics simulations (GROMACS) to assess binding stability (RMSD < 2Å)
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Dock antibody-antigen complexes using HADDOCK2.4 with interface residues from homolog studies
Validation pipeline:
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In silico affinity maturation (FoldX)
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Deep mutational scanning of CDR regions
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High-throughput SPR screening (10<sup>4</sup> variants/day)
