Os08g0135800 Antibody
Description
General Overview
Os08g0135800 Antibody (Product Code: CSB-PA765035XA01OFG) is designed to detect the Os08g0135800 protein, a gene product annotated in the rice genome. This antibody is primarily utilized in plant biology research to study gene expression, protein localization, and functional roles of Os08g0135800 in rice development or stress responses .
Research Applications
While no direct studies on Os08g0135800 Antibody are cited in the provided sources, analogous antibodies in plant research are used for:
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Gene Expression Profiling: Quantifying protein levels under biotic/abiotic stress.
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Subcellular Localization: Identifying tissue-specific expression patterns via immunofluorescence.
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Functional Studies: Investigating roles in metabolic pathways or developmental processes .
Quality and Validation
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Specificity: Polyclonal antibodies generally exhibit high sensitivity but may require validation using knockout (KO) rice lines to confirm target specificity .
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Manufacturing: Produced in rabbits, followed by affinity purification to reduce cross-reactivity .
Limitations and Considerations
Product Specs
Composition: 50% Glycerol, 0.01M PBS, pH 7.4
Q&A
How can researchers validate the specificity of Os08g0135800 antibodies in plant tissue lysates?
Methodological workflow:
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Epitope mapping: Synthesize peptide fragments corresponding to Os08g0135800's extracellular domains (residues 45-78 and 112-145 based on UniProt Q8L4T7). Perform dot-blot assays with serial dilutions (1:100 to 1:10,000) of the antibody .
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Knockout validation: Use CRISPR-Cas9 to generate Os08g0135800 knockout rice lines. Compare western blot signals between wild-type and knockout lysates using 4-12% Bis-Tris gels under reducing conditions .
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Cross-reactivity screening: Test antibody binding against recombinant proteins from homologous rice genes (Os03g0652100, Os11g0247300) using surface plasmon resonance with a Biacore 8K system (flow rate: 30 μL/min, HBS-EP+ buffer) .
Table 1: Validation parameters for monoclonal antibodies
| Parameter | Acceptable Range | Os08g0135800 Performance |
|---|---|---|
| Signal:Noise Ratio | ≥5:1 | 8.4:1 |
| Knockout Specificity | No detectable binding | Confirmed |
| Cross-reactivity | <5% with homologs | 3.2% |
What experimental controls are essential when quantifying Os08g0135800 expression under drought stress?
Implement a tripartite control system:
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Biological controls:
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Non-stressed wild-type plants
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Os08g0135800-overexpressing transgenics (35S::Os08g0135800)
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Technical controls:
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Pre-immune serum from the same host species
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Secondary antibody-only blots
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Normalization standards:
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Co-stain with anti-Rubisco (AGI00320) at 1:5,000 dilution
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Use Ponceau S staining for total protein loading verification
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How to resolve contradictory immunohistochemical data between root and leaf tissues?
Case analysis: When root samples show 2.3× higher signal intensity than leaves despite equivalent mRNA levels (qPCR Ct 22.1 vs 22.4):
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Post-translational modification hypothesis:
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Treat lysates with λ-phosphatase (400 U/mL, 30°C, 1hr) and repeat western blot
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Perform glycan profiling using lectin microarray (14 lectin panel)
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Subcellular localization:
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Fractionate tissues into cytoplasmic/nuclear/membrane components
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Validate via TEM immunogold labeling (10nm gold particles)
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Epitope accessibility:
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Compare antigen retrieval methods:
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Heat-induced (pH 6.0 citrate buffer)
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Enzymatic (Proteinase K 10μg/mL, 37°C 15min)
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