At3g17490 Antibody
Description
Introduction to At3g17490 Antibody
The At3g17490 Antibody is a targeted immunological reagent designed to detect proteins encoded by the At3g17490 gene in Arabidopsis thaliana (thale cress). It is a custom antibody produced by Cusabio, a biotechnology company specializing in antibody development. This antibody is primarily used in plant molecular biology research to study protein localization, function, and interactions, particularly in contexts involving F-box domain-containing proteins and ubiquitination pathways .
Gene Function and Antibody Utility
The At3g17490 gene encodes a protein containing an F-box domain, a motif critical for interactions with Skp1/Cullin/F-box (SCF) E3 ubiquitin ligase complexes. These complexes mediate the ubiquitination and degradation of target proteins, regulating processes such as cell cycle control, hormone signaling, and stress responses in plants .
Potential Research Focus Areas:
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Protein Degradation Studies:
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Subcellular Localization:
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Determining the intracellular localization of the target protein (e.g., nucleus, cytoplasm).
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Interaction Mapping:
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Identifying binding partners using co-immunoprecipitation (Co-IP) or proximity-based assays.
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Antibody Performance
While specific experimental data for the At3g17490 Antibody is limited in public literature, Cusabio’s product listings emphasize its suitability for standard immunological techniques. For example:
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Western Blot: Detecting the target protein in Arabidopsis lysates.
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Immunofluorescence: Visualizing protein localization in plant tissues .
Comparison with Related Antibodies:
Optimal Experimental Conditions
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Dilution: Typically 1:1000–1:2000 for Western blotting, depending on sample complexity .
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Blocking Buffer: 5% milk or BSA to minimize nonspecific binding .
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Detection: ECL (enhanced chemiluminescence) or fluorescent secondary antibodies .
Limitations and Challenges
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Species Specificity: Limited to Arabidopsis, restricting cross-species studies .
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Antibody Availability: Exclusively distributed by Cusabio, with no open-source alternatives reported .
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Functional Validation: No published neutralization or inhibition assays for this antibody, unlike therapeutic antibodies (e.g., bimekizumab ).
Product Specs
Constituents: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Q&A
FAQs for Researchers on At3g17490 Antibody
While no direct data on At3g17490 antibody exists in the provided search results, the following FAQs are constructed based on general antibody research methodologies and best practices in academic settings. These reflect common challenges and analytical frameworks applicable to plant protein antibodies like At3g17490 (a Arabidopsis thaliana gene product).
What experimental controls are critical for immunohistochemistry (IHC) with At3g17490 antibody?
Advanced Considerations:
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Negative Tissue Controls: Use tissues from At3g17490 knockout plants.
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Isotype Controls: Substitute primary antibody with same-host species non-specific IgG.
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Fixation Optimization: Test paraformaldehyde concentrations (2–4%) to balance epitope preservation and tissue integrity.
How to resolve discrepancies in At3g17490 antibody performance across laboratories?
Data Contradiction Analysis Framework:
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Technical Variability: Compare protocols (e.g., fixation time, antibody dilution).
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Biological Variability: Assess plant growth conditions (light, temperature) affecting protein expression.
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Lot-to-Lot Variation: Request antibody validation data from suppliers for different batches.
How to optimize At3g17490 antibody for co-immunoprecipitation (Co-IP) assays?
Optimization Steps:
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Crosslinker Choice: Test non-cleavable (e.g., DSS) vs. cleavable (e.g., DTSSP) reagents.
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Elution Buffer: Compare low-pH (glycine buffer) vs. competitive peptide elution.
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Post-IP Validation: Confirm interactions via Western blot or mass spectrometry.
What statistical approaches are recommended for low-signal At3g17490 antibody data?
Advanced Analysis:
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Background Subtraction: Use rolling-ball algorithm in imaging software.
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Replicate Stratification: Pool data from ≥3 biological replicates to mitigate plant-to-plant variability.
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Signal Amplification: Employ tyramide-based amplification for low-abundance targets.
How to design a CRISPR-based validation system for At3g17490 antibody specificity?
Experimental Design:
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Generate CRISPR-Cas9 At3g17490 knockouts in Arabidopsis.
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Introduce silent mutations in the epitope region (preserving protein function but altering antibody binding).
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Compare antibody binding between wild-type, knockout, and epitope-edited lines.
Notes for Researchers:
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At3g17490 is implicated in [hypothetical function based on gene annotation], necessitating rigorous validation in stress-response or developmental studies.
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Collaborate with plant biology consortia (e.g., TAIR) for shared validation protocols.
