At1g05670 Antibody
Description
Introduction to At1g05670 Antibody
The At1g05670 Antibody is a specialized immunological reagent designed to target the protein encoded by the AT1G05670 gene in Arabidopsis thaliana (thale cress). This antibody enables precise detection and study of the protein’s subcellular localization, functional interactions, and regulatory roles in plant molecular biology.
Subcellular Localization Studies
The At1g05670 Antibody has been used to investigate the protein’s localization in Arabidopsis. Fluorescence microscopy studies show:
This dual targeting suggests the protein may regulate RNA processes in both organelles .
Role in Cytokinin Signaling
In ethylene and cytokinin pathway studies, AT1G05670 was identified alongside response regulators (e.g., ARR2) and cytokinin-related genes (e.g., AT2G40670) . While direct functional evidence is limited, its association with these pathways implies potential involvement in growth regulation or stress responses.
Challenges and Future Directions
While the antibody’s role in basic research is established, gaps remain:
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Functional Mechanism: Direct evidence linking AT1G05670 to RNA processing or signaling pathways is lacking.
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Species-Specificity: Limited data on cross-reactivity with non-Arabidopsis organisms.
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Therapeutic Potential: Unlike monoclonal antibodies in veterinary medicine (e.g., Solensia for osteoarthritis) , this antibody’s translational applications remain unexplored.
Product Specs
Composition: 50% Glycerol, 0.01M Phosphate Buffered Saline (PBS), pH 7.4
Target Background
Q&A
What is At1g05670 Antibody and what is its target?
At1g05670 Antibody is a polyclonal antibody raised in rabbits against recombinant Arabidopsis thaliana At1g05670 protein. The antibody recognizes the native protein encoded by the At1g05670 gene in Arabidopsis thaliana (Mouse-ear cress), a model organism widely used in plant molecular biology. The antibody is generated through antigen affinity purification and is designed for research applications only .
What are the key specifications of commercially available At1g05670 Antibody?
The antibody has the following specifications:
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Product Type: Polyclonal Antibody
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Host Organism: Rabbit
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Target: Recombinant At1g05670 protein (Arabidopsis thaliana)
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Applications: ELISA, Western Blot
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Form: Liquid
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Storage Buffer: 0.03% Proclin 300, 50% Glycerol, 0.01M PBS, pH 7.4
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UniProt Number: Q0WVK7
| Property | Specification |
|---|---|
| Product Code | CSB-PA605890XA01DOA |
| Immunogen | Recombinant Arabidopsis thaliana At1g05670 protein |
| Isotype | IgG |
| Clonality | Polyclonal |
| Purification Method | Antigen Affinity Purified |
| Lead Time | Made-to-order (14-16 weeks) |
How should researchers optimize Western blotting protocols for At1g05670 Antibody?
While specific manufacturer protocols may vary, researchers should consider the following optimization strategy for Western blotting with At1g05670 Antibody:
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Sample preparation: Extract plant proteins using appropriate lysis buffers containing protease inhibitors. Consider using specialized plant protein extraction buffers that account for cell wall components and phenolic compounds.
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Dilution optimization: Test a range of antibody dilutions to determine optimal signal-to-noise ratio. The table below provides guidance based on typical polyclonal antibody performance:
| Antibody Dilution | Signal Strength | Background | Signal-to-Noise Ratio |
|---|---|---|---|
| 1:500 | Strong | High | Moderate |
| 1:1000 | Strong | Moderate | Good |
| 1:2000 | Moderate | Low | Excellent |
| 1:5000 | Weak | Very Low | Moderate |
| 1:10000 | Very Weak | Very Low | Poor |
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Blocking optimization: For plant proteins, test both conventional (5% milk/BSA) and plant-specific blocking agents to reduce background.
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Incubation conditions: For primary antibody, test both overnight at 4°C and 2-4 hours at room temperature to determine optimal conditions.
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Detection method selection: Choose chemiluminescence for highest sensitivity or colorimetric methods for easier quantification.
What controls are essential when using At1g05670 Antibody in experiments?
When performing experiments with At1g05670 Antibody, include the following controls:
Positive controls:
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Wild-type Arabidopsis thaliana tissue extracts
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Recombinant At1g05670 protein (if available)
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Tissue known to express high levels of the target protein
Negative controls:
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At1g05670 knockout mutant tissue extracts
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Primary antibody omission
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Blocking peptide competition assay
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Non-expressing tissues or developmental stages
Procedural controls:
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Loading control antibodies (anti-actin or anti-tubulin)
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Molecular weight marker
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Non-specific rabbit IgG at equivalent concentration
What are the most common technical issues when using At1g05670 Antibody and how can they be resolved?
Common issues and their solutions include:
| Issue | Potential Causes | Solutions |
|---|---|---|
| No signal | Insufficient antibody concentration, protein degradation, incorrect secondary antibody | Increase antibody concentration, add fresh protease inhibitors, verify secondary antibody compatibility |
| High background | Insufficient blocking, excessive antibody concentration, inadequate washing | Increase blocking time, reduce antibody concentration, increase wash steps |
| Multiple bands | Cross-reactivity, protein degradation, post-translational modifications | Pre-absorb antibody, add protease inhibitors, use phosphatase inhibitors if studying phosphorylated forms |
| Inconsistent results | Antibody degradation, sample variation | Aliquot antibody, standardize protein extraction method |
| Weak signal | Low protein expression, inefficient transfer | Increase protein loading, optimize transfer conditions |
How can researchers verify the specificity of At1g05670 Antibody?
To verify antibody specificity:
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Genetic validation: Compare immunoblots using wild-type versus At1g05670 knockout/knockdown plant tissues.
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Peptide competition: Pre-incubate antibody with excess immunizing peptide before application to block specific binding sites.
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Molecular weight confirmation: Verify that the detected band corresponds to the predicted molecular weight of At1g05670 protein.
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Immunoprecipitation-Mass Spectrometry: Perform immunoprecipitation followed by mass spectrometry to confirm target identity.
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Heterologous expression: Test antibody against heterologous expression systems (e.g., bacteria, yeast) expressing recombinant At1g05670.
How can At1g05670 Antibody be used for protein-protein interaction studies?
For protein-protein interaction studies:
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Co-Immunoprecipitation (Co-IP): Use At1g05670 Antibody to pull down the target protein and identify interacting partners by Western blot or mass spectrometry.
| Buffer Type | Advantages | Disadvantages | Recommended For |
|---|---|---|---|
| RIPA | Good solubilization | May disrupt weak interactions | Strong interactions |
| NP-40 (0.5%) | Preserves interactions | Less efficient extraction | Sensitive protein complexes |
| Plant-specific buffers | Optimized for plant tissues | May require optimization | Initial screening |
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Proximity Ligation Assay (PLA): Combine At1g05670 Antibody with antibodies against potential interacting proteins to visualize interactions in situ.
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FRET Analysis: Use fluorescently labeled secondary antibodies against At1g05670 Antibody and potential interactors.
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Pull-down validation: Complement antibody-based interaction studies with recombinant protein pull-downs.
What experimental approaches can be used to study At1g05670 localization in plant tissues?
To study protein localization:
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Immunohistochemistry protocol optimization:
| Parameter | Variables to Test | Optimization Notes |
|---|---|---|
| Fixation | 4% PFA, Methanol, Acetone | Test different fixatives for best epitope preservation |
| Antigen retrieval | Heat-induced, Enzymatic | May be necessary for formalin-fixed tissues |
| Antibody dilution | 1:100 to 1:500 | Start with higher concentration for IHC than WB |
| Incubation time | 1-3 hours RT vs. overnight 4°C | Longer incubations often yield better results |
| Detection system | Fluorescent vs. Chromogenic | Choose based on required sensitivity and imaging facilities |
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Subcellular fractionation followed by immunoblotting: Separate cellular compartments (nucleus, cytoplasm, membrane, chloroplast) and detect At1g05670 in each fraction.
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Immunogold electron microscopy: For high-resolution localization studies at the ultrastructural level.
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Correlative light and electron microscopy (CLEM): Combine immunofluorescence with electron microscopy for comprehensive localization analysis.
How can At1g05670 Antibody be used in plant stress response studies?
For investigating stress responses:
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Expression profiling: Monitor At1g05670 protein levels under various stress conditions (drought, salt, pathogen, heat) using quantitative Western blot.
| Stress Condition | Sample Collection Timing | Special Considerations |
|---|---|---|
| Drought | Early, mid, and late stress | Control water loss carefully |
| Salt stress | 1h, 6h, 24h post-treatment | Use consistent NaCl concentrations |
| Cold stress | During and after cold exposure | Process samples without thawing |
| Heat stress | During heat shock and recovery | Prevent protein degradation during collection |
| Pathogen infection | Early (6-12h) and late (24-48h) | Include mock-infected controls |
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PTM analysis: Investigate stress-induced post-translational modifications by combining immunoprecipitation with PTM-specific antibodies.
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Protein stability studies: Determine if stress affects protein turnover rate using cycloheximide chase assays followed by immunoblotting.
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Chromatin immunoprecipitation (ChIP): If At1g05670 is a DNA-binding protein, examine stress-induced changes in DNA binding using ChIP-qPCR or ChIP-seq.
What complementary techniques should be used alongside At1g05670 Antibody to achieve comprehensive results?
For comprehensive analysis:
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Transcriptional analysis: Combine protein detection with RT-qPCR or RNA-seq to correlate transcript and protein levels.
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Transgenic approaches: Use overexpression or CRISPR/Cas9 knockout lines to validate antibody specificity and protein function.
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Protein-metabolite interactions: Couple immunoprecipitation with metabolomics to identify associated metabolites.
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Systems biology integration: Contextualize At1g05670 protein data within broader protein-protein interaction networks, metabolic pathways, or signaling cascades.
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Proteomic profiling: Combine with quantitative proteomics to study changes in the entire proteome when At1g05670 expression is altered.
How might At1g05670 Antibody contribute to understanding plant adaptation mechanisms?
The At1g05670 Antibody can facilitate research into plant adaptation through:
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Monitoring protein expression across different ecotypes grown under various environmental conditions
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Investigating protein modifications in response to climate change-related stresses
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Studying protein-protein interaction networks that may reveal adaptation mechanisms
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Examining evolutionary conservation of At1g05670 function across related species
What emerging technologies could enhance the utility of At1g05670 Antibody in research?
Emerging technologies include:
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Single-cell proteomics: Using At1g05670 Antibody for high-resolution cellular analysis
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Multi-omics integration: Combining antibody-based protein data with transcriptomics, metabolomics, and phenomics
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Spatial proteomics: Applying the antibody in tissue-clearing methods for whole-plant imaging
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Microfluidic antibody arrays: High-throughput analysis of At1g05670 across multiple conditions simultaneously
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AI-assisted image analysis: Enhancing quantification of immunolocalization results
