At3g57590 Antibody
Description
Western Blotting (WB)
The antibody demonstrates high specificity in WB, enabling detection of At3g57590 in Arabidopsis protein extracts. Protocols typically involve:
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Dilution: 1:1000 starting concentration.
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Detection: Compatible with standard chemiluminescent or colorimetric systems.
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Validation: Dot blot assays confirm detection limits of 0.01–1 ng of immunogen peptide .
ELISA and Immunoprecipitation (IP)
While primarily optimized for WB, the antibody is also suitable for ELISA-based antigen-antibody interaction studies. For IP, the AbInsure™ program covers experimental validation, though specific protocols require optimization .
Research Relevance and Functional Insights
The At3g57590 F-box protein belongs to the SCF (SKP1-CUL1-F-box) complex, which mediates ubiquitination-dependent protein degradation in plants. While no direct studies on this antibody’s use in functional assays are publicly documented, its utility lies in:
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Protein Localization: Tracking At3g57590 expression in Arabidopsis tissues.
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Pathway Analysis: Studying interactions with other SCF complex components.
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Stress Response: Investigating the protein’s role in environmental stress adaptation (e.g., temperature, salinity).
Cross-Reactivity and Specificity
Tissue cross-reactivity studies (as outlined in ) are critical for minimizing off-target binding. For plant-specific antibodies, species-specific validation is essential to avoid false positives from analogous proteins in other organisms.
Stability and Aggregation
Therapeutic antibodies often face aggregation challenges, which can be assessed via mass photometry . While not explicitly tested for At3g57590, protocols for forced degradation (e.g., pH, temperature stress) could be adapted to evaluate its stability.
Antibody Design and Target Structure
The N-terminal targeting strategy aligns with common epitope selection for F-box proteins, which often lack conserved structural motifs. Structural databases like AbDb or PLAbDab could theoretically map the antibody’s binding site, though no crystallographic data are available for At3g57590.
| Feature | Detail |
|---|---|
| Target Class | F-box protein (involved in E3 ubiquitin ligase activity) |
| Epitope | N-terminal synthetic peptide |
| Orthologs | Limited to Arabidopsis based on current data |
Availability and Cost
The antibody is marketed as part of the X2 package ($899) or X3.AT5G14740.5 (not detailed here), with shipping and handling costs ($100). Delivery typically occurs within 30 days .
Product Specs
Q&A
Given the specific nature of the query about "At3g57590 Antibody," which does not appear in the provided search results, I will create a general FAQ collection for researchers focusing on antibody-related research, particularly in academic scenarios. This will cover aspects such as experimental design, data analysis, and methodological considerations.
A:
To validate the specificity of an antibody, you should:
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Use multiple controls: Include both positive and negative controls to ensure the antibody binds specifically to the target antigen.
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Perform Western blotting: This technique helps confirm the antibody's specificity by detecting the target protein in a complex mixture.
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Conduct immunoprecipitation (IP) assays: IP can help verify the antibody's ability to bind and precipitate the target protein specifically.
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Use orthogonal methods: Techniques like mass spectrometry or other biochemical assays can further validate the findings.
A:
When encountering contradictory results:
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Re-evaluate experimental conditions: Ensure that conditions such as temperature, buffer composition, and incubation times are consistent across experiments.
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Check antibody specificity: Verify that the antibody is specific to the target antigen using techniques like Western blot or IP.
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Consider batch variability: If using different batches of the same antibody, test both batches under identical conditions to rule out batch-specific effects.
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Use independent validation methods: Employ alternative detection methods (e.g., qPCR for mRNA levels) to confirm protein expression or localization.
A:
Antibody subclasses (e.g., IgG1 vs. IgG3) can influence therapeutic efficacy due to differences in effector functions and half-life. For instance, IgG3 has a longer hinge region, potentially enhancing flexibility and effector functions, but its reduced plasma half-life and challenges in recombinant expression limit its use . Understanding these differences is crucial for designing effective therapeutic antibodies.
A:
To optimize antibody binding conditions in SPR assays:
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Buffer selection: Use a buffer that minimizes non-specific binding and maintains antibody stability.
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Temperature control: Ensure consistent temperature conditions as this can affect binding kinetics.
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Flow rate adjustment: Optimize the flow rate to achieve the best binding signal without compromising the sensor chip.
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Regeneration conditions: Use appropriate regeneration solutions to remove bound antibodies without damaging the sensor chip.
A:
Several computational tools are available for antibody design, such as RosettaAntibodyDesign (RAbD) and IgDesign. These tools allow for the design of antibody complementarity-determining regions (CDRs) to enhance binding affinity and specificity . They can predict and optimize antibody structures based on antigen-antibody complexes, facilitating the creation of high-affinity antibodies.
Example Data Table: Comparison of Antibody Design Tools
| Tool | Key Features | Application Area |
|---|---|---|
| RAbD | Samples diverse sequence and structure space | Broad range of applications |
| IgDesign | Uses deep learning for CDR design | Therapeutic antigen binding |
These tools are essential for advancing antibody research by enabling the design of antibodies with tailored properties for specific applications.
