At4g15060 Antibody
Description
Introduction to At4g15060 Antibody
The At4g15060 Antibody is a research-grade monoclonal antibody targeting the protein encoded by the At4g15060 gene in Arabidopsis thaliana (mouse-ear cress), a model organism in plant biology. This antibody is commercially available through specialized biotechnology suppliers (e.g., Cusabio) and is utilized to study gene expression, protein localization, and functional roles of the At4g15060 protein in cellular processes .
Gene Expression Studies
The At4g15060 Antibody has been employed to analyze gene expression patterns in Arabidopsis. For example, in a study investigating histone demethylation in Arabidopsis, the antibody was used to detect downregulated expression of At4g15060 under specific experimental conditions:
| Experiment | Observation |
|---|---|
| Gene Expression | At4g15060 expression ↓ (p = 0.0541) |
| Antibody Dilution | 1:170 |
| Context | Linked to epigenetic regulation |
This suggests the antibody’s role in studying chromatin remodeling or stress responses in plants .
Target Protein Context
The At4g15060 gene encodes a protein potentially involved in F-box protein-related pathways, based on homology with known F-box proteins in Arabidopsis. These proteins often participate in ubiquitin-mediated protein degradation, a key regulatory mechanism in cellular processes .
Antibody Specificity and Validation
While no peer-reviewed validation studies for this specific antibody are available in the provided sources, its use in Western blot and immunohistochemistry implies compatibility with standard protocols for plant protein detection. Commercial suppliers typically validate antibodies against recombinant proteins or knockout (KO) samples to ensure specificity .
Limitations in Current Data
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Limited Public Data: Most information about the At4g15060 Antibody comes from commercial catalogs, with sparse experimental details in peer-reviewed literature.
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Species-Specificity: The antibody is validated for Arabidopsis, limiting cross-reactivity claims with other plant species.
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Dilution Optimization: While a 1:170 dilution was effective in one study, optimal conditions may vary by application (e.g., immunoprecipitation vs. IHC) .
Research Gaps
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Mechanistic Insights: The biological role of the At4g15060 protein remains poorly characterized.
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Functional Studies: No data exist on the antibody’s utility in functional assays (e.g., protein interaction studies).
Product Specs
Composition: 50% Glycerol, 0.01M PBS, pH 7.4
Q&A
Basic Research Questions
How to validate specificity of At4g15060 antibody in Arabidopsis protein extracts?
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Perform Western blot with knockout mutants (e.g., T-DNA insertion lines) to confirm absence of signal in negative controls .
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Combine immunofluorescence with transgenic lines expressing GFP-tagged At4g15060 for subcellular localization cross-validation .
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Use peptide competition assays by pre-incubating antibodies with excess antigenic peptides (10:1 molar ratio) .
What experimental controls are critical for ChIP-seq using At4g15060 antibody?
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Include:
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IgG isotype controls for nonspecific binding
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Input DNA normalization
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Mutant tissue lacking At4g15060 as biological negative control
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Validate ≥3 biological replicates using qPCR for known target loci before sequencing .
How to address inconsistent Western blot signals across developmental stages?
Advanced Research Questions
How to resolve contradictory co-IP/MS data involving At4g15060?
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Perform reciprocal immunoprecipitations with tag-specific antibodies
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Apply crosslinking (1% formaldehyde, 10 min) before lysis to preserve transient interactions
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Analyze raw MS data with SAINTexpress (FDR <0.01) and exclude contaminants via CRAPome database
What strategies enable single-cell resolution studies of At4g15060?
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Optimize CITE-seq protocols using titrated antibody concentrations (0.025-0.62 µg/mL) :
Cell Type Optimal Conc. Sequencing Saturation Root protoplasts 0.15 µg/mL 85% Leaf mesophyll 0.25 µg/mL 78% -
Pair with 10x Genomics Multiome ATAC + antibody oligo conjugation
How to map conformational epitopes recognized by At4g15060 antibody?
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Generate alanine-scanning mutants across O23360 (UniProt)
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Express mutant proteins in Nicotiana benthamiana via agroinfiltration
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Quantify antibody binding affinity using BLI (Octet RED96):
Target: M for structural studies
