At3g62790 Antibody

What is At3g62790 and what role does it play in plant biology?

At3g62790 (UniProt: Q9LZI6) is a protein found in Arabidopsis thaliana that functions as part of Complex I in the mitochondrial respiratory chain. This protein is involved in electron transport and energy metabolism in plant cells, making it an important subject for research in plant stress responses and metabolic pathways .

Methodological approach to studying At3g62790 function:

• Comparative genomic analysis across different plant species

• Phenotypic characterization of At3g62790 knockouts or overexpression lines

• Subcellular localization studies using fluorescent protein fusions

• Analysis of expression patterns under various environmental conditions

How are At3g62790 antibodies properly validated for research use?

Validation of At3g62790 antibodies requires multiple complementary approaches to ensure specificity and reliability:

Standard validation protocol:

• Western blot analysis - Confirm antibody detects a protein of the expected molecular weight (~75 kDa) in Arabidopsis extracts

• Negative controls - Test reactivity against extracts from At3g62790 knockout mutants

• Immunoprecipitation followed by mass spectrometry - Verify the identity of the pulled-down protein

• Cross-reactivity testing - Evaluate potential cross-reactivity with related proteins in the same family

Validation data example:

When validating polyclonal antibodies like CSB-PA878646XA01DOA, additional batch-to-batch consistency testing is essential .

What are the optimal storage and handling conditions for At3g62790 antibodies?

Proper storage and handling of At3g62790 antibodies is critical for maintaining their functionality:

Storage conditions:

• Store at -20°C or -80°C upon receipt

• Avoid repeated freeze-thaw cycles by preparing small working aliquots

• Most commercial At3g62790 antibodies are supplied in a stabilizing buffer containing 50% glycerol and 0.03% Proclin 300 as a preservative

Handling recommendations:

• Thaw aliquots completely before use and mix gently (do not vortex)

• Centrifuge briefly before opening vials to collect liquid at the bottom

• For working dilutions, use freshly prepared buffer (typically PBS with 1% BSA)

• Document lot numbers, receipt dates, and freezer locations in laboratory records

The half-life of properly stored antibodies can exceed 5 years, but functionality should be confirmed periodically through positive control experiments.

What applications are At3g62790 antibodies suitable for in plant research?

At3g62790 antibodies have been validated for several laboratory techniques:

Confirmed applications:

• Western blotting (WB): For detecting denatured At3g62790 protein in plant extracts

• Enzyme-linked immunosorbent assay (ELISA): For quantitative measurement of At3g62790 levels

• Immunohistochemistry (IHC): For localization studies in fixed plant tissues

Experimental considerations by application:

These applications make At3g62790 antibodies valuable tools for studying protein expression patterns, especially in the context of plant stress responses .

What experimental design considerations are critical for At3g62790 antibody-based studies?

Robust experimental design is essential for obtaining reliable results with At3g62790 antibodies:

Key experimental design elements:

• Statistical power analysis: Determine appropriate sample size and replication based on expected effect size

• Randomization: Arrange samples randomly to avoid systematic bias

• Appropriate controls: Include both positive and negative controls in each experiment

• Blocking factors: Control for variables that might affect results (e.g., plant age, growth conditions)

• Blinding: When possible, blind the researcher to sample identity during analysis

Advanced design considerations:

• Split-plot design: When testing multiple variables (e.g., genotype × stress × time)

• Latin square design: To control for position effects in plate-based assays

• Factorial design: To efficiently test multiple factors and their interactions

Example experimental matrix for At3g62790 stress response study:

Proper experimental design significantly increases the statistical power to detect true effects while minimizing false positives .

How can epitope mapping improve At3g62790 antibody specificity and application?

Epitope mapping is crucial for understanding antibody binding characteristics and optimizing experimental protocols:

Epitope mapping methodologies:

• Peptide array analysis: Synthesize overlapping peptides spanning the At3g62790 sequence and test antibody binding

• Mutational analysis: Introduce point mutations to identify critical binding residues

• Hydrogen-deuterium exchange mass spectrometry (HDX-MS): Identify regions of the protein protected by antibody binding

• X-ray crystallography: Determine the three-dimensional structure of the antibody-antigen complex

Benefits of epitope knowledge:

• Enables prediction of cross-reactivity with homologous proteins

• Guides optimization of sample preparation methods (e.g., fixation conditions)

• Informs development of blocking peptides for specificity controls

• Facilitates selection of appropriate antibody combinations for multiplexing

Knowledge of the specific epitope recognized by an At3g62790 antibody is particularly important when studying related plant proteins or when extending research to other plant species .

How can machine learning approaches enhance At3g62790 antibody development and application?

Machine learning is revolutionizing antibody research through several innovative approaches:

Machine learning applications in antibody research:

• Epitope prediction: Algorithms can predict likely epitopes based on protein sequence and structure

• Cross-reactivity prediction: Models can identify potential off-target binding

• Optimization of experimental conditions: Machine learning can identify optimal parameters for antibody-based assays

• Active learning for binding prediction: Reducing experimental costs by intelligently selecting which experiments to perform

Implementation strategies:

• Transfer learning: Apply models trained on general antibody-antigen interactions to At3g62790-specific applications

• Ensemble methods: Combine multiple predictive models to improve accuracy

• Bayesian optimization: Systematically identify optimal experimental conditions

Recent advances in machine learning for antibody-antigen binding prediction have shown up to 35% reduction in required experimental testing through active learning approaches , which could significantly accelerate At3g62790 antibody research.

What troubleshooting strategies should be employed when At3g62790 antibodies perform suboptimally?

Systematic troubleshooting is essential when facing challenges with At3g62790 antibody applications:

Common issues and solutions matrix:

Methodological approach to troubleshooting:

• Test antibody with positive control sample (recombinant protein)

• Systematically adjust one variable at a time

• Document all modifications to protocols

• Validate findings with alternative detection methods

This structured approach can identify whether issues originate from the antibody itself, the experimental conditions, or the biological sample .

What are the latest advances in generating high-affinity At3g62790 antibodies using recombinant antibody technology?

Recombinant antibody technology offers significant advantages for developing improved At3g62790 antibodies:

Advanced antibody engineering approaches:

• Phage display libraries: Selection of high-affinity antibody fragments

• Yeast surface display: Engineering antibodies with improved specificity

• Deep mutational scanning: Systematic testing of antibody variants

• Inverse folding models: Computational design of antibody complementarity-determining regions (CDRs)

Benefits of recombinant antibodies for At3g62790 research:

• Consistent renewable source without batch-to-batch variation

• Precise control over antibody characteristics

• Ability to introduce specific modifications (e.g., fusion tags)

• Reduced dependency on animals for antibody production

Recent advances in antibody engineering have demonstrated successful design of high-affinity binders with up to 83% success rates using inverse folding models , suggesting potential for developing improved At3g62790 antibodies with enhanced specificity and affinity.

How can At3g62790 antibodies contribute to understanding plant adaptation mechanisms to environmental stress?

At3g62790 antibodies can provide critical insights into plant adaptation mechanisms:

Research applications for environmental adaptation studies:

• Comparative analysis: Track At3g62790 protein levels across ecotypes from different environments

• Field-to-lab translation: Compare protein expression in controlled conditions versus field experiments

• Transgenic studies: Analyze phenotypic consequences of At3g62790 modification

• Signaling pathway elucidation: Map At3g62790 interactions in stress response networks

Experimental approach for adaptation studies:

• Collect diverse ecotypes from varying environments

• Expose to controlled stress conditions (e.g., drought, temperature, aluminum)

• Measure At3g62790 protein levels using validated antibodies

• Correlate expression patterns with stress tolerance phenotypes

• Identify regulatory mechanisms through genetic and protein interaction studies

This approach can reveal how At3g62790 contributes to adaptive responses, similar to studies on ALS3 and PGIP1 in Arabidopsis, which demonstrated their roles in aluminum stress responses through expression-GWAS approaches .