ACR11 Antibody
Description
ACR11 Antibody Overview
The ACR11 antibody targets the ACT domain-containing protein ACR11 (AT1G16880), which plays a critical role in nitrogen metabolism by regulating the stability and activity of ferredoxin-dependent glutamate synthase (Fd-GOGAT). Key features include:
Research Applications
The ACR11 antibody has been employed in diverse experimental approaches to elucidate ACR11's function:
Protein Interaction Studies
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Fd-GOGAT Complex Formation: Co-immunoprecipitation (Co-IP) and Blue Native PAGE (BN-PAGE) revealed that ACR11 forms an ~800 kDa protein complex with Fd-GOGAT, essential for maintaining Fd-GOGAT stability and activity. Loss of ACR11 in acr11 mutants disrupts this complex, leading to reduced Fd-GOGAT protein levels and activity .
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Post-Translational Regulation: Immunoblot analyses demonstrated that ACR11 stabilizes Fd-GOGAT post-transcriptionally. In acr11 mutants, Fd-GOGAT protein levels drop by 50–70%, while mRNA levels remain unchanged .
Response to Metabolic Changes
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Diurnal Regulation: ACR11 mediates diurnal fluctuations in Fd-GOGAT levels, with protein amounts peaking during the day and declining at night. This rhythm is absent in acr11 mutants .
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Nitrogen (N) Signaling: Upon N resupply, ACR11 enables rapid recovery of Fd-GOGAT levels in wild-type plants, a response absent in acr11 mutants despite comparable glutamine accumulation .
Key Research Findings
The table below summarizes critical discoveries enabled by the ACR11 antibody:
Functional Implications
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Nitrogen Metabolism: ACR11 ensures efficient glutamate synthesis by stabilizing Fd-GOGAT, a key enzyme in the GS/GOGAT cycle. This regulation is vital for adapting to N availability and photorespiration .
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Pathogen Resistance: acr11 mutants show elevated reactive oxygen species (ROS) and salicylic acid (SA), linking ACR11 to redox balance and defense responses .
Product Specs
Constituents: 50% Glycerol, 0.01M PBS, pH 7.4
Target Background
Q&A
Given the constraints and the focus on academic research scenarios, I have compiled a list of FAQs related to the ACR11 Antibody that reflect the depth of scientific research and methodological considerations. These questions are designed to address both basic and advanced research scenarios, focusing on experimental design, data analysis, and methodological approaches.
Answer:
To study the role of ACR11 in plant metabolism, researchers can employ a combination of biochemical and molecular biology techniques. This includes:
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Immunoblotting: Use the ACR11 antibody to detect changes in ACR11 protein levels in different plant tissues or under varying conditions.
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Co-immunoprecipitation: Identify interacting proteins by using the ACR11 antibody to precipitate ACR11 and associated proteins, followed by mass spectrometry analysis.
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Mutant Analysis: Compare metabolic profiles between wild-type plants and ACR11 mutants to understand its functional role.
Answer:
Discrepancies can arise due to differences in sensitivity, specificity, or sample preparation. To resolve these:
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Validate Antibodies: Ensure the specificity of the ACR11 antibody by testing it against known positive and negative controls.
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Standardize Protocols: Use consistent sample preparation and experimental conditions across methods.
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Quantitative Analysis: Use quantitative techniques like mass spectrometry to validate qualitative findings from immunoblotting.
Answer:
To study protein-protein interactions involving ACR11:
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Blue Native PAGE (BN-PAGE): Separate protein complexes based on their native size and charge, followed by immunoblotting with the ACR11 antibody.
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Co-immunoprecipitation (Co-IP): Use the ACR11 antibody to pull down ACR11 and associated proteins, which can then be identified by mass spectrometry.
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Proximity Ligation Assay (PLA): Visualize protein interactions in situ using fluorescence microscopy.
Answer:
Validation involves:
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Knockout or Knockdown Controls: Test the antibody against tissues or cells lacking ACR11 expression.
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Western Blot and Immunoblotting: Verify the antibody's ability to detect ACR11 specifically in different plant extracts.
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Immunoprecipitation Followed by Mass Spectrometry: Confirm that the antibody specifically pulls down ACR11 and not other proteins.
Answer:
Integration involves:
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Multidisciplinary Approaches: Combine biochemical, molecular, and physiological data to understand ACR11's function.
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Bioinformatics Tools: Use databases and software to analyze protein interactions and metabolic pathways involving ACR11.
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Statistical Analysis: Apply statistical methods to identify significant patterns or correlations in the data.
Answer:
Advanced techniques include:
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RNA Interference (RNAi): Knockdown ACR11 expression to study its effects on metabolism.
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CRISPR-Cas9 Gene Editing: Create ACR11 mutants for detailed functional analysis.
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Proteomics and Metabolomics: Use mass spectrometry-based approaches to analyze changes in protein and metabolite profiles in response to ACR11 modulation.
Answer:
Challenges include:
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Antibody Specificity: Validate antibodies rigorously to ensure specificity.
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Sample Preparation: Standardize sample preparation to minimize variability.
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Data Interpretation: Use multiple approaches to validate findings and address discrepancies.
Answer:
Future directions include:
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Exploring ACR11's Role in Stress Responses: Investigate how ACR11 modulates plant metabolism under stress conditions.
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Elucidating the Mechanism of ACR11-Mediated Regulation: Use biochemical and structural biology techniques to understand how ACR11 interacts with other proteins and regulates metabolic pathways.
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Applying CRISPR-Cas9 for Crop Improvement: Use gene editing to modify ACR11 expression in crops to enhance metabolic traits beneficial for agriculture.
